Publishing type：Research paper (scientific journal)  

The classical organ culture method, in which tissue is placed at the gas‒liquid interphase, is effective at inducing mouse spermatogenesis. However, due to reginal variations in the supply of oxygen and nutrients within a tissue, the progress of spermatogenesis was observed only in limited areas of a tissue. In addition, haploid cell formation and its differentiation to spermatozoon, i.e. spermiogenesis, were infrequent and inefficient. Here, we show that the polydimethylsiloxane (PDMS)-chip ceiling (PC) method, which ensures a uniform supply of nutrients and oxygen throughout the tissue by pressing it into a thin, flat shape, can provide control over the culture space. We used this method to culture testis tissue from neonatal mice, aged 1 to 4 days, and found that modulating the culture space during the experiment by replacing one chip with another that had a higher ceiling effectively increased tissue growth. This adjustment also induced more efficient spermatogenesis, with the process of spermiogenesis being particularly promoted. Meiotic cells were observed from culture day 14 onward, and haploid cells were confirmed at the end of each experiment. This technique was also shown to be a sensitive assay for testicular toxicity. Culture-space control will be a critical regulation parameter for sophisticated tissue culture experiments.

DOI： 10.1038/s41598-023-39323-y

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PORTFOLIO  

An in vitro spermatogenesis method using mouse testicular tissue to produce fertile sperm was established more than a decade ago. Although this culture method has generally not been effective in other animal species, we recently succeeded in improving the culture condition to induce spermatogenesis of rats up to the round spermatid stage. In the present study, we introduced acrosin-EGFP transgenic rats in order to clearly monitor the production of haploid cells during spermatogenesis in vitro. In addition, a metabolomic analysis of the culture media during cultivation revealed the metabolic dynamics of the testis tissue. By modifying the culture media based on these results, we were able to induce rat spermatogenesis repeatedly up to haploid cell production, including the formation of elongating spermatids, which was confirmed histologically and immunohistochemically. Finally, we performed a microinsemination experiment with in vitro produced spermatids, which resulted in the production of healthy and fertile offspring. This is the first demonstration of the in vitro production of functional haploid cells that yielded offspring in animals other than mice. These results are expected to provide a basis for the development of an in vitro spermatogenesis system applicable to many other mammals.

DOI： 10.1038/s41598-023-39304-1

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Publishing type：Research paper (scientific journal)   Publisher：Public Library of Science (PLoS)  

Mouse spermatogenesis, from spermatogonial stem cell proliferation to sperm formation, can be reproduced in vitro by culturing testis tissue masses of neonatal mice. However, it remains to be determined whether this method is also applicable when testis tissues are further divided into tiny fragments, such as segments of the seminiferous tubule (ST), a minimal anatomical unit for spermatogenesis. In this study, we investigated this issue using the testis of an Acrosin-GFP/Histone H3.3-mCherry (Acr/H3) double-transgenic mouse and monitored the expression of GFP and mCherry as indicators of spermatogenic progression. Initially, we noticed that the cut and isolated stretches of ST shrunk rapidly and conglomerated. We therefore maintained the isolation of STs in two ways: segmental isolation without truncation or embedding in soft agarose. In both cases, GFP expression was observed by fluorescence microscopy. By whole-mount immunochemical staining, meiotic spermatocytes and round and elongating spermatids were identified as Sycp3-, crescent-form GFP-, and mCherry-positive cells, respectively. Although the efficiency was significantly lower than that with tissue mass culture, we clearly showed that spermatogenesis can be induced up to the elongating spermatid stage even when the STs were cut into short segments and cultured in isolation. In addition, we demonstrated that lowered oxygen tension was favorable for spermatogenesis both for meiotic progression and for producing elongating spermatids in isolated STs. Culturing isolated STs rather than tissue masses is advantageous for explicitly assessing the various environmental parameters that influence the progression of spermatogenesis.

DOI： 10.1371/journal.pone.0283773

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Language：Japanese   Publisher：(株)ニュー・サイエンス社  

不妊症の約50%に男性側因子が関与している。精巣内精子回収法(Testicular Sperm Extraction:TESE)や卵細胞質内精子注入法(Intracytoplasmic Sperm Injection:ICSI)の普及で,無精子症患者の挙児も可能になった。しかし男性不妊症の主要因である精子形成不全の病態解明は今後の課題である。近年の生殖細胞生物学の発展を鑑み,近未来の男性不妊症治療の展開を予想する。(著者抄録)

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Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Reproduction  

DOI： 10.1262/jrd.2023-093

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Language：Japanese   Publisher：医学図書出版(株)  

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Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

Abstract

Mammalian spermatogenesis is a heat-vulnerable process that occurs at low temperatures, and elevated testicular temperatures cause male infertility. However, the current reliance on in vivo assays limits their potential to detail temperature dependence and destructive processes. Using ex vivo cultures of mouse testis explants at different controlled temperatures, we found that spermatogenesis failed at multiple steps, showing sharp temperature dependencies. At 38 °C (body core temperature), meiotic prophase I is damaged, showing increased DNA double-strand breaks (DSBs) and compromised DSB repair. Such damaged spermatocytes cause asynapsis between homologous chromosomes and are eliminated by apoptosis at the meiotic checkpoint. At 37 °C, some spermatocytes survive to the late pachytene stage, retaining high levels of unrepaired DSBs but do not complete meiosis with compromised crossover formation. These findings provide insight into the mechanisms and significance of heat vulnerability in mammalian spermatogenesis.

DOI： 10.1038/s42003-022-03449-y

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Other Link： https://www.nature.com/articles/s42003-022-03449-y

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Mammalian male germ-cell development consists of three distinct phases: primordial germ cell (PGC) development, male germ-cell specification for spermatogonium development, and ensuing spermatogenesis. Here, we show an in vitro reconstitution of whole male germ-cell development by pluripotent stem cells (PSCs). Mouse embryonic stem cells (mESCs) are induced into PGC-like cells (mPGCLCs), which are expanded for epigenetic reprogramming. In reconstituted testes under an optimized condition, such mPGCLCs differentiate into spermatogonium-like cells with proper developmental transitions, gene expression, and cell-cycle dynamics and are expanded robustly as germline stem cell-like cells (GSCLCs) with an appropriate androgenetic epigenome. Importantly, GSCLCs show vigorous spermatogenesis, not only upon transplantation into testes in vivo but also under an in vitro culture of testis transplants, and the resultant spermatids contribute to fertile offspring. By uniting faithful recapitulations of the three phases of male germ-cell development, our study creates a paradigm for the in vitro male gametogenesis by PSCs.

DOI： 10.1016/j.stem.2021.08.005

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)  

© 2021, The Author(s). In vitro spermatogenesis (IVS) using air–liquid interphase organ culture method is possible with mouse testis tissues. The same method, however, has been hardly applicable to animals other than mice, only producing no or limited progression of spermatogenesis. In the present study, we challenged IVS of rats with modifications of culture medium, by supplementing chemical substances, including hormones, antioxidants, and lysophospholipids. In addition, reducing oxygen tension by placing tissues in an incubator of lower oxygen concentration and/or applying silicone cover ceiling on top of the tissue were effective for improving the spermatogenic efficiency. Through these modifications of the culture condition, rat spermatogenesis up to round spermatids was maintained over 70 days in the cultured tissue. Present results demonstrated a significant progress in rat IVS, revealing conditions commonly favorable for mice and rats as well as finding rat-specific optimizations. This is an important step towards successful IVS in many animal species, including humans.

DOI： 10.1038/s41598-021-82792-2

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

<title>Abstract</title>
Spermatogenesis takes place in the seminiferous tubules, starting from the spermatogonial stem cell and maturing into sperm through multiple stages of cell differentiation. Sertoli cells, the main somatic cell constituting the seminiferous tubule, are in close contact with every germ cell and play pivotal roles in the progression of spermatogenesis. In this study, we developed an in vitro Sertoli cell replacement method by combining an organ culture technique and a toxin receptor-mediated cell knockout system. We used Amh-diphtheria toxin receptor transgenic mice, whose Sertoli cells specifically express human diphtheria toxin receptor, which renders them sensitive to diphtheria toxin. An immature Amh-diphtheria toxin receptor testis was transplanted with the donor testis cells followed by culturing in a medium containing diphtheria toxin. This procedure successfully replaced the original Sertoli cells with the transplanted Sertoli cells, and spermatogenesis originating from resident germ cells was confirmed. In addition, Sertoli cells in the mouse testis tissues were replaced by transplanted rat Sertoli cells within culture conditions without requiring immunosuppressive treatments. This method works as a functional assay system, making it possible to evaluate any cells that might function as Sertoli cells. It would also be possible to investigate interactions between Sertoli and germ cells more closely, providing a new platform for the study of spermatogenesis and its impairments.

DOI： 10.1093/biolre/ioab104

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Other Link： http://academic.oup.com/biolreprod/article-pdf/105/4/934/40545414/ioab104.pdf

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PURPOSE: To assess the effect of our new classification on surgical outcomes after flexible ureteroscopy (fURS) for kidney stones. METHODS: We retrospectively examined 128 patients after single renal fURS procedures performed using ureteral access sheaths (UASs) with the fragmentation technique. Based on the gap (calculated by subtracting the ureteroscope diameter from the UAS diameter), enrolled patients were divided into three groups: small (< 0.6 mm), medium (0.6 to < 1.2 mm), and large space groups (≥ 1.2 mm). Stone-free (SF) status was defined as either complete absence of stones (SF) or the presence of stones < 4 mm in diameter on non-contrast computed tomography (NCCT). RESULTS: The SF rate was significantly lower in the small space group (50% in small, 97.9% in medium, 89.2% in large; p = 0.001). Perioperative complications over Clavien-Dindo Grade I were observed in 16.7%, 4.2%, and 8.1% of patients, respectively (p = 0.452). The ratio of stone volume and operative time (efficiency of stone removal) was significantly higher in the large space group compared to the small and medium space groups (0.009 ± 0.003 ml/min, 0.013 ± 0.005 ml/min, 0.027 ± 0.012 ml/min, respectively; p < 0.001). CONCLUSION: Our findings that gaps > 0.6 mm (1.8 Fr), including the combination of a 9.5-Fr UAS and a small caliber ureteroscope, improve SF rates, and larger gaps facilitate stone removal efficiency providing the basis for future development of clinical protocols aimed at improving outcomes.

DOI： 10.1007/s00345-020-03459-7

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

In vitro spermatogenesis, which produces fertile spermatozoa, has been successfully performed using an organ culture method from murine tissue. Here, we provide a dataset of time-course microarray transcriptome data of in vitro cultured neonate murine testes and age-matched in vivo-derived testes. The dataset presented here is related to the article titled "Transcriptome analysis reveals inadequate spermatogenesis and immediate radical immune reactions during organ culture in vitro spermatogenesis" published in Biochemical and Biophysical Research Communications in 2020 [1]. The raw data and pre-processed data are publicly available on the GEO repository (accession number GSE147982). Furthermore, the dataset provided here includes additional metadata, detailed explanations of the experiment, results of pre-processing, analysis scripts, and lists of differentially expressed genes from in vitro culture testes and in vivo testes at each time point.

DOI： 10.1016/j.dib.2020.106482

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Language：Japanese   Publisher：(有)科学評論社  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Cultivation of neonatal mouse testis tissue can induce spermatogenesis and produce fertile sperms. However, in vitro spermatogenesis mediated by the current organ culture method comes short in fully mimicking the in vivo counterpart, partly due to a lack of knowledge underlying molecular phenotypes of in vitro spermatogenesis. In this study, we investigated transcriptome of cultured testis tissues using microarray method. Principle component analysis of the transcriptome data revealed delay and/or arrest of spermatogenesis and immediate radical immune reactions in the cultured testis tissues. The delay/arrest of spermatogenesis occurred before and during early meiotic phase, resulting in inefficient progression of meiosis. The immune reaction, on the other hand, was drastic and overwhelming, in which TLR4-NF-kB signaling was speculated to be involved. Notably, treatment with TAK242, an inhibitor of TLR4-NF-kB signaling pathway, ameliorated the macrophage activation which otherwise would exacerbate the inflammation. Thus, the present study revealed for the first time at molecular level that the deficiency of germ cell differentiation and the immense immune reaction are major abnormalities in the cultured testis tissues.

DOI： 10.1016/j.bbrc.2020.06.161

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Language：English   Publishing type：Research paper (scientific journal)  

PURPOSE: To identify risk factors by developing and internally validating a nomogram for preventing perioperative complications in overnight ureteral catheterization cases after fURS for kidney stones. METHODS: We retrospectively examined 309 patients with overnight ureteral catheterization after single fURS procedures for renal stones. fURS procedures were performed based on the fragmentation technique. The ureteral catheter was removed on postoperative day 1. Within this group, patients who experienced perioperative complications (complication group) were compared with those who did not experience complications (non-complication group). The complication group included 77 patients whose Clavien-Dindo classification score was I, II, III, or IV and/or those whose body temperature during hospitalization was over 37.5 °C. RESULTS: The overall stone volume, stone-free rate, incidence of perioperative complications, and procedure duration were 1.39 mL, 94.8%, 24.9%, and 62 min, respectively. Severe complications of a Clavien-Dindo level III or IV were observed in only four cases (1.3%). Multivariate assessment revealed five independent predictors of perioperative complications after fURS with overnight catheterization: age (p = 0.11), sex (p = 0.067), stone volume (p = 0.33), Hounsfield units (p = 0.16), and narrow ureter (p = 0.018). We developed a nomogram to predict perioperative complications after fURS using these parameters. CONCLUSIONS: We developed a predictive model for perioperative complications of patients with overnight catheterization after fURS for renal stones. This model could select patients who were at a low risk of complications.

DOI： 10.1007/s00345-019-03023-y

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Publishing type：Research paper (scientific journal)   Publisher：Radiation Research Society  

DOI： 10.1667/rade-19-00018.1

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© 2020 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology In vitro mouse spermatogenesis using a classical organ culture method became possible by supplementing basal culture medium with only the product of bovine serum albumin purified by chromatography (AlbuMAX), which indicated that AlbuMAX contained every chemical factor necessary for mouse spermatogenesis. However, since the identity of these factors was unclear, improvements in culture media and our understanding of the nutritional and signal substances required for spermatogenesis were hindered. In the present study, chemically defined media (CDM) without AlbuMAX was used to evaluate each supplementary factor and their combinations for the induction of spermatogenesis. Similar to in vivo conditions, retinoic acid, triiodothyronine (T3), and testosterone (T) were needed. Based on differences in spermatogenic competence between AlbuMAX, fetal bovine serum, and adult bovine serum, we identified α-tocopherol, which strongly promoted spermatogenesis when combined with ascorbic acid and glutathione. Differences were also observed in the abilities of lipids extracted from AlbuMAX using two different methods to induce spermatogenesis. This led to the identification of lysophospholipids, particularly lysophosphatidylcholine, lysophosphatidic acid, and lysophosphatidylserine, as important molecules for spermatogenesis. New CDM formulated based on these results induced and promoted spermatogenesis as efficiently as AlbuMAX-containing medium. In vitro spermatogenesis with CDM may provide a unique experimental system for research on spermatogenesis that cannot be performed in in vivo experiments.

DOI： 10.1096/fj.202000245R

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PURPOSE: The effect of combining miniaturization with endoscopic combined intrarenal surgery (ECIRS) is unclear. Thus, we compared the treatment outcomes between minimally invasive ECIRS (mini-ECIRS) using 16.5 Fr percutaneous access sheath and standard ECIRS using 24 Fr access sheath for renal stones MATERIALS AND METHODS: We retrospectively analyzed consecutive patients who underwent single session mini or standard-ECIRS in the modified Valdivia position for renal stones between April 2009 and May 2016. To adjust for patient characteristics, 77 pairs were matched using preoperative parameters including age, sex, history of febrile urinary tract infection (UTI), stone surface area, number of involved calyces, and staghorn calculi. RESULTS: The stone free rate (SFR) was similar between mini and standard ECIRS according to non-contrast computed tomography (61.1% vs. 52.0%, p = 0.388). The rate of perioperative complications exceeding grade 2 based on the Clavien-Dindo classification was similar in both groups (19.5% vs. 26.0%, p = 0.442). Severe complications exceeding grade 3 were also similar in both groups (2.6% vs. 3.9%, p > 0.99). Two cases of septic shock were noted in each group. Although there was no difference regarding bleeding-related complications (2.6% vs. 6.5%, p = 0.442), pseudoaneurysm or blood transfusion was not observed in the mini-ECIRS group. Pain visual analog scale values in the perioperative period were lower in the mini-ECIRS group (1.34 ± 1.08 vs. 1.69 ± 1.23, p = 0.062). CONCLUSIONS: This study demonstrated that, compared to standard ECIRS, mini-ECIRS maintained SFR without increasing perioperative complications, tended to reduce postoperative pain and had a potential to reduce bleeding-related complications. This report suggests the advantages of ECIRS miniaturization for renal stones.

DOI： 10.1007/s11255-020-02433-x

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Language：English  

Radiotherapy can result in temporary or permanent gonadal toxicity in male cancer patients despite the high precision and accuracy of modern radiation treatment techniques. Previous radiobiological studies have shown an effective tissue-sparing response in various tissue types and species following exposure to spatially fractionated radiation. In the present study, we used an ex vivo mouse testicular tissue culture model and a conventional X-ray irradiation device to evaluate the tissue-sparing effect (TSE) of spatially fractionated X-rays for the protection of male fertility from radiotherapy-related adverse effects. We revealed a significant TSE for maintaining spermatogenesis in the ex vivo testes model following spatially fractionated X-ray irradiation. Moreover, we experimentally propose a possible mechanism by which the migration of spermatogonial cells, from the non-irradiated areas to the irradiated ones, in irradiated testicular tissue, is essential for the TSE and maintaining spermatogenesis. Therefore, our findings demonstrate that the control of TSE following spatially fractionated X-rays in the testes has a considerable potential for clinical application. Interdisciplinary research will be essential for further expanding the applicability of this method as an approach for the preservation of male fertility during or after radiotherapy.

DOI： 10.3390/jcm9041089

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Language：Japanese   Publisher：(一社)日本内分泌学会  

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Language：Japanese   Publisher：(一社)日本内分泌学会  

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Language：English   Publishing type：Research paper (scientific journal)  

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：Japanese   Publisher：医学図書出版(株)  

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：Japanese   Publisher：(一社)日本病理学会  

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Language：English   Publishing type：Research paper (scientific journal)  

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Language：Japanese   Publisher：(一社)日本内分泌学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

DOI： 10.1002/bit.26822

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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DOI： 10.1063/1.5035328

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier B.V.  

In our previous study, we produced a microfluidic device (MFD) which successfully maintained spermatogenesis for over 6 months in mouse testis tissues loaded in the device. In the present study, we developed a new MFD, a monolayer device (ML-D) with a barrier structure consisting of pillars and slits, which is simpler in design and easier to make. This ML-D was also effective for inducing mouse spermatogenesis and maintained it for a longer period than the conventional culture method. In addition, we devised a way of introducing sample tissue into the device during its production, just before bonding the upper layer of polydimethylsiloxane (PDMS) and bottom glass slide. The tissue can obtain nutrients horizontally from the medium running beside it and oxygen vertically from above through PDMS. In addition, the glass slide set at the bottom improved the visibility of the sample tissue with an inverted microscope. When we took photos of cultured tissue of the Acr-Gfp transgenic mouse testis in ML-D sequentially every day, morphological changes of the acrosome during spermiogenesis were successfully recorded. The ML-D is simple in design and useful for culturing testis tissue for inducing and maintaining spermatogenesis with clearer visibility. Due to the new method of sample loading, tissues other than testis should also be applicable.

DOI： 10.1016/j.bbrc.2018.04.180

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DOI： 10.1667/RR14957.1

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DOI： 10.1007/s00345-018-2328-1

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Public Library of Science  

We previously reported the successful induction and completion of mouse spermatogenesis by culturing neonatal testis tissues. The culture medium consisted of α-minimum essential medium (α-MEM), supplemented with Knockout serum replacement (KSR) or AlbuMAX, neither of which were defined chemically. In this study, we formulated a chemically defined medium (CDM) that can induce mouse spermatogenesis under organ culture conditions. It was found that bovine serum albumin (BSA) purified through three different procedures had different effects on spermatogenesis. We also confirmed that retinoic acid (RA) played crucial roles in the onset of spermatogonial differentiation and meiotic initiation. The added lipids exhibited weak promoting effects on spermatogenesis. Lastly, luteinizing hormone (LH), follicle stimulating hormone (FSH), triiodothyronine (T3), and testosterone (T) combined together promoted spermatogenesis until round spermatid production. The CDM, however, was not able to produce elongated spermatids. It was also unable to induce spermatogenesis from the very early neonatal period, before 2 days postpartum, leaving certain factors necessary for spermatogenic induction in mice unidentified. Nonetheless, the present study provided important basic information on testis organ culture and spermatogenesis in vitro.

DOI： 10.1371/journal.pone.0192884

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1080/09553002.2017.1355579

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Three-dimensional aggregation and organ culture methods are critical for recreating in vivo cellular phenomena outside the body. Previously, we used the conventional gas liquid interphase organ culture method to induce complete mouse spermatogenesis. After incorporating microfluidic systems, we achieved a significant increase in efficiency and duration of spermatogenesis. One of the major drawbacks preventing the popularization of microfluidics, however, is the use of a power-pump to generate medium flow. In this study, we produced a pumpless microfluidic device using hydrostatic pressure and a resistance circuit to facilitate slow, longer lasting medium flow. During three months of culture, results in induction and maintenance of spermatogenesis showed no difference between pumpless and pump-driven devices. Correspondingly, the spermatogonial population was favorably maintained in the pumpless device compared to the conventional method. These results show the advantage of using microfluidic systems for organ culture experiments. Our pumpless device could be applied to a variety of other tissues and organs, and may revolutionize organ culture methods as a whole.

DOI： 10.1038/s41598-017-15799-3

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

A search for early response genes that are activated following germ cell induction from mouse embryonic stem cells in vitro led us to the isolation of a long noncoding RNA that contains a SINE (short interspersed element)-B1F motif that was named R53. In situ hybridization and northern blot analyses revealed that the R53 subfragment RNA bears a B1F motif, is processed from the primary transcript, is expressed in adult testis and is predominantly localized in meiotic metaphase chromatin during spermatogenesis. Recent studies of chromosome-associated RNAs have explored novel functions of noncoding RNAs. Specifically, chromosome-bound noncoding RNAs function not only as structural components of chromosome but also as scaffolds that recruit epigenetic modulators for transcriptional regulation, and they are dynamically rearranged during the cell cycle. However, few studies have explored meiotic chromatin; thus, R53 RNA appears to be the first long noncoding RNA to be tightly associated with the metaphase chromatin during spermatogenesis. Furthermore, R53 knockdown using a lentivirus-mediated RNAi injected into mouse testis and organ culture of the fragments revealed a remarkable reduction in postmeiotic cells and irregular up-regulation of several postmeiotic genes, which suggests the possibility that the SINE-B1-derived noncoding RNA R53 plays an indispensable role in the transcriptional regulation of key spermatogenesis genes.

DOI： 10.1371/journal.pone.0179585

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

Background: The complexity of spermatogenesis makes development of appropriate in vitro testis models challenging. A novel in vitro mouse testis culture system has been reported but not yet evaluated as an alternative model for male reproductive toxicity testing. We assessed the effects of media composition on sperm differentiation and testis morphology of cultured mouse testis fragments. Methods: Testes from postnatal day 5 B6: CBA-Tg(Acrv1-EGFP) 2727Redd/J male mice were cultured in knockout serum replacement (KSR) or Albumax I (Albumax) medium. Enhanced green fluorescent protein (EGFP) expression was examined on days 35, 42, 45, and 49 of culture. Histology and flow cytometry were performed for testis morphology and spermatid differentiation. Results: EGFP signals were first observed in round spermatids on day 22 of culture (corresponding to postnatal day 27) and were observed until the end of culture, indicating testis-specific protein expression. A-kinase anchor protein 4 expression, a marker of elongated spermatid (step 15-16) occurred earlier in explants cultured in KSR than Albumax medium (typically day 35 and after day 42 of culture, respectively). The percentage of seminiferous tubules with elongated spermatid was higher in Albumax than KSR medium from days 45 to 49 of culture. Conclusion: Albumax medium may facilitate or support better morphology and spermatid production than KSR medium. Further studies need to improve spermatid production and refinement of this in vitro testis culture system that may be useful as a supplement to current male reproductive toxicity testing or an alternative model in cases where in vivo testing may be unfeasible. (C) 2017 Wiley Periodicals, Inc.

DOI： 10.1002/bdr2.1002

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：CELL PRESS  

In the seminiferous tubules of mouse testes, a population of glial cell line-derived neurotrophic factor family receptor alpha 1 (GFR alpha 1)positive spermatogonia harbors the stem cell functionality and supports continual spermatogenesis, likely independent of asymmetric division or definitive niche control. Here, we show that activation of Wnt/beta-catenin signaling promotes spermatogonial differentiation and reduces the GFR alpha 1(+) cell pool. We further discovered that SHISA6 is a cell-autonomous Wnt inhibitor that is expressed in a restricted subset of GFRa1(+) cells and confers resistance to the Wnt/b-catenin signaling. Shisa6(+) cells appear to show stem cell-related characteristics, conjectured from the morphology and long-term fates of T (Brachyury)(+) cells that are found largely overlapped with Shisa6(+) cells. This study proposes a generic mechanism of stem cell regulation in a facultative (or open) niche environment, with which different levels of a cell-autonomous inhibitor (SHISA6, in this case) generates heterogeneous resistance to widely distributed differentiation-promoting extracellular signaling, such as WNTs.

DOI： 10.1016/j.stemcr.2017.01.006

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

We previously reported the successful induction of complete spermatogenesis of mice in neonatal testis tissues cultured on agarose gel, with the culture medium supplemented with a bovine serum albumin product, AlbuMAX. This method, however, has not been examined for fetal testis tissues. In this report, we tested the culture method for fetal testes of the Acrosin (Acr)-Gfp transgenic mouse, whose testicular germ cells express GFP from the midmeiotic phase onward, using Albu-MAX-containing medium. The fetal testis, from 19.5 days postcoitum (dpc) back to 14.5 dpc, showed spermatogenic progression and produced haploid cells in culture. On the other hand, testes of 13.5 dpc or earlier did not show the meiotic sign of Acr-Gfp expression. Regardless of the fetal age, tissue masses enlarged during the culture period because of the elongation and thickening of the seminiferous tubules. This simple culture method could be a useful experimental system to investigate fetal testicular development and germ cell biology.

DOI： 10.1095/biolreprod.116.140277

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Language：Japanese   Publisher：日本生殖内分泌学会  

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J-GLOBAL

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Language：Japanese   Publisher：(一社)日本抗加齢医学会  

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

In contrast to cell cultures, particularly to cell lines, tissues or organs removed from the body cannot be maintained for long in any culture conditions. Although it is apparent that in vivo regional homeostasis is facilitated by the microvascular system, mimicking such a system ex vivo is difficult and has not been proved effective. Using the culture system of mouse spermatogenesis, we addressed this issue and devised a simple microfluidic device in which a porous membrane separates a tissue from the flowing medium, conceptually imitating the in vivo relationship between the microvascular flow and surrounding tissue. Testis tissues cultured in this device successfully maintained spermatogenesis for 6 months. The produced sperm were functional to generate healthy offspring with micro-insemination. In addition, the tissue kept producing testosterone and responded to stimulation by luteinizing hormone. These data suggest that the microfluidic device successfully created in vivo-like conditions, in which testis tissue maintained its physiologic functions and homeostasis. The present model of the device, therefore, would provide a valuable foundation of future improvement of culture conditions for various tissues and organs, and revolutionize the organ culture method as a whole.

DOI： 10.1038/srep21472

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：John Wiley and Sons Ltd  

Cancer treatments, either chemo- or radiotherapy, may cause severe damage to gonads which could lead to the infertility of patients. In post-pubertal male patients, semen cryopreservation is recommended to preserve the potential to have their own biological children in the future
however, it is not applicable to prepubertals. The preservation of testis tissue which contains spermatogonial stem cells (SSCs) but not sperm would be an alternative measure. The tissues or SSCs have to be transplanted back into patients to obtain sperm
however, this procedure remains experimental, invasive, and is accompanied with the potential risk of re-implantation of cancer cells. Recently, we developed an organ culture system which supports the spermatogenesis of mice up to sperm formation from SSCs. It was also shown that the tissues could be frozen for later sperm production, which resulted in the generation of offspring. Thus, it could be useful as a clinical application for preserving the reproductive potential of male pediatric cancer patients. The establishment of an optimized cryopreservation method and the development of a culture system for human testis tissue are expected in the future.

DOI： 10.1007/s12522-015-0218-4

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Language：English   Publisher：MEDKNOW PUBLICATIONS & MEDIA PVT LTD  

Twenty years ago, the transplantation of spermatogonial stem cells (SSCs) from a mouse to other recipient mice was shown to be feasible, which clearly demonstrated the functional identity of SSCs. Since then, several important new findings and other technical developments have followed, which included a new hypothesis on their cell kinetics and spermatogonial hierarchy in the testis, a culture method allowing their self-renewal and proliferation, a testis tissue organ culture method, which induced their complete differentiation up to sperm, and the in vitro induction of germ cells from embryonic stem cells and induced pluripotent stem cells. These advancements reinforced or advanced our understanding of this unique cell. Nonetheless, there are many unresolved questions in the study of spermatogonial stem cells and a long road remains until these cells can be used clinically in reproductive medicine.

DOI： 10.4103/1008-682X.154995

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Language：Japanese   Publisher：(一社)日本内分泌学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：CELL PRESS  

Mouse spermatogonial stem cells (SSCs) can be cultured for multiplication and maintained for long periods while preserving their spermatogenic ability. Although the cultured SSCs, named germline stem (GS) cells, are targets of genome modification, this process remains technically difficult. In the present study, we tested TALEN and double-nicking CRISPR/Cas9 on GS cells, targeting Rosa26 and Stra8 loci as representative genes dispensable and indispensable in spermatogenesis, respectively. Harvested GS cell colonies showed a high targeting efficiency with both TALEN and CRISPR/Cas9. The Rosa26-targetedGS cells differentiated into fertility-competent sperm following transplantation. On the other hand, Stra8-targeted GS cells showed defective spermatogenesis following transplantation, confirming its prime role in the initiation of meiosis. TALEN and CRISPR/Cas9, when applied in GS cells, will be valuable tools in the study of spermatogenesis and for revealing the genetic mechanism of spermatogenic failure.

DOI： 10.1016/j.stemcr.2015.05.011

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Language：Japanese   Publisher：(有)科学評論社  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

Research on in vitro spermatogenesis is important for elucidating the spermatogenic mechanism. We previously developed an organ culture method which can support spermatogenesis from spermatogonial stem cells up to sperm formation using immature mouse testis tissues. In this study, we examined whether it is also applicable to mature testis tissues of adult mice. We used two lines of transgenic mice, Acrosin-GFP and Gsg2-GFP, which carry the marker GFP gene specific for meiotic and haploid cells, respectively. Testis tissue fragments of adult GFP mice, aged from 4 to 29 weeks old, which express GFP at full extension, were cultured in medium supplemented with 10% KSR or AlbuMAX. GFP expression decreased rapidly and became the lowest at 7 to 14 days of culture, but then slightly increased during the following culture period. This increase reflected de novo spermatogenesis, confirmed by BrdU labeling in spermatocytes and spermatids. We also used vitamin A-deficient mice, whose testes contain only spermatogonia. The testes of those mice at 13-21 weeks old, showing no GFP expression at explantation, gained GFP expression during culturing, and spermatogenesis was confirmed histologically. In addition, the adult testis tissues of Sl/Sl(d) mutant mice, which lack spermatogenesis due to Kit ligand mutation, were cultured with recombinant Kit ligand to induce spermatogenesis up to haploid formation. Although the efficiency of spermatogenesis was lower than that of pup, present results showed that the organ culture method is effective for the culturing of mature adult mouse testis tissue, demonstrated by the induction of spermatogenesis from spermatogonia to haploid cells.

DOI： 10.1371/journal.pone.0130171

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：Japanese   Publisher：医学図書出版(株)  

前立腺肥大症と診断した50歳以上、IPSS(International Prostate SymptomScore:国際前立腺症状スコア)8点以上かつQOLスコア4点以上の21症例を対象とした。ナフトピジル75mg/日を経口投与し、投与後1、2、3および14日目でのIPSS、QOLスコア、OABSSを投与前と比較し評価した。総IPSSは投与後2日目で有意なスコアの低下を認め、さらに投与後14日目で最大効果発現を示した。IPSS蓄尿症状スコアにおいても同様の傾向が認められた。安全性については、21例中1例に動悸が出現したが、症状は軽度であり継続投与可能であった。ナフトピジル75mg/日は早期、特に2日目から症状の改善を認め、前立腺肥大症に伴う自覚症状に対し有用であった。また、その安全性も確認された。(著者抄録)

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

With the increasing cure rate of paediatric cancers, infertility, as one of the adverse effects of treatments, has become an important concern for patients and their families. Since semen cryopreservation is applicable only for post-pubertal patients, alternative pre-pubertal measures are necessary. Here we demonstrate that testis tissue cryopreservation is a realistic measure for preserving the fertility of an individual. Testis tissues of neonatal mice were cryopreserved either by slow freezing or by vitrification. After thawing, they were cultured on agarose gel and showed spermatogenesis up to sperm formation. Microinsemination was performed with round spermatids and sperm, leading to eight offspring in total. They grew healthily and produced progeny upon natural mating between them. This strategy, the cryopreservation of testis tissues followed by in vitro spermatogenesis, is promising to preserve the fertility of male paediatric cancer patients in the future.

DOI： 10.1038/ncomms5320

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Language：Japanese   Publisher：(株)メディカルレビュー社  

精子幹細胞は精子形成の根幹を成しており、その分化増殖メカニズムの研究は男性不妊の病態解明と治療法の開発に必要不可欠である。近年、精子幹細胞の研究は大きく進展しており、マウスでは器官培養法を用いて精子幹細胞からの精子産生がin vitroで可能となった。さらにES細胞やiPS細胞から精子を産生することが可能になった。今後ヒトへの応用が可能となれば、男性不妊の研究の進展が期待される。(著者抄録)

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Language：Japanese   Publisher：(一社)日本泌尿器科学会総会事務局  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

We investigated the Core Lower Urinary Tract Symptom Score as an outcome assessment tool for the treatment of lower urinary tract symptoms using silodosin. In addition, the ability of the Core Lower Urinary Tract Symptom Score to detect overactive bladder in male patients with lower urinary tract symptoms was examined. The present study included 241 males with benign prostatic hyperplasia treated at 31 medical facilities between June 2009 and December 2010. All patients were given silodosin, and the effects of silodosin intake were measured using four questionnaires: the Core Lower Urinary Tract Symptom Score, International Prostate Symptom Score, Overactive Bladder Symptom Score and Quality-of-Life index. The efficacy of silodosin for treating lower urinary tract symptoms was validated according to the total scores of all four questionnaires weighted equally (P&lt;0.05). Spearman's among the Core Lower Urinary Tract Symptom Score, International Prostate Symptom Score and Overactive Bladder Symptom Score showed a mild-high correlation. However, the correlation between the baseline values of the Core Lower Urinary Tract Symptom Score and Quality-of-Life index was low in the groups with benign prostatic hyperplasia (=0.314) and benign prostatic hyperplasia/overactive bladder (=0.244). Our findings showed the Core Lower Urinary Tract Symptom Score, both its total score and each subscore, is able to show the efficacy of silodosin, similar to other questionnaires. The Core Lower Urinary Tract Symptom Score is also useful for identifying overactive bladder symptoms in patients with benign prostatic hyperplasia. As the Core Lower Urinary Tract Symptom Score does not correlate well with the Quality-of-Life index, these two questionnaires might be better used in combination to assess treatment outcomes.

DOI： 10.1111/iju.12167

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

The in vitro propagation of mouse spermatogonial stem cells (SSCs) became possible in 2003; these cultured SSCs were named germ-line stem (GS) cells. To date, however, it has not been possible to induce spermatogenesis from GS cells in vitro. Recently, we succeeded in producing functional sperm from primitive spermatogonia in explanted neonatal mouse testis tissues. Here we describe a protocol that can support spermatogenesis from GS cells up to sperm formation in vitro using an organ culture method. GS cells transplanted in the extracted testis form colonies in the tissue fragments and differentiate into sperm under the described in vitro organ culture conditions. It takes about 6 weeks to obtain sperm from GS cells. The sperm are viable, resulting in healthy offspring through micro-insemination. Thus, this protocol should be a valuable tool for the study of mammalian spermatogenesis.

DOI： 10.1038/nprot.2013.138

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：Japanese   Publisher：(一社)日本癌治療学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

Cells of testicular tissues during fetal or neonatal periods have the ability to reconstruct the testicular architecture even after dissociation into single cells. This ability, however, has not been demonstrated effectively in vitro. In the present study, we reconstructed seminiferous tubules in vitro that supported spermatogenesis to the meiotic phase. First, testicular cells of neonatal mice were dissociated enzymatically into single cells. Then, the cells formed aggregates in suspension culture and were transferred to the surface of agarose gel to continue the culture with a gas-liquid interphase method, and a tubular architecture gradually developed over the following 2 wk. Immunohistological examination confirmed Sertoli cells forming tubules and germ cells inside. With testicular tissues of Acr-GFP transgenic mice, the germ cells of which express GFP during meiosis, cell aggregates formed a tubular structure and showed GFP expression in their reconstructed tissues. Meiotic figures were also confirmed by regular histology and immunohistochemistry. In addition, we mixed cell lines of spermatogonial stem cells (GS cells) into the testicular cell suspension and found the incorporation of GS cells in the tubules of reconstructed tissues. When GS cells derived from Acr-GFP transgenic mice were used, GFP expression was observed, indicating that the spermatogenesis of GS cells was proceeding up to the meiotic phase. This in vitro reconstruction technique will be a useful method for the study of testicular organogenesis and spermatogenesis.

DOI： 10.1095/biolreprod.113.108613

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Language：Japanese   Publisher：日本小児泌尿器科学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARY ANN LIEBERT, INC  

Purpose: To investigate the utility and limitations of stone surface area (SA) as a predictor of stone-free (SF) status after a single semirigid ureteroscopy (URS), with or without a flexible component, for the treatment of patients with urinary stones.
Patients and Methods: Cases of patients with urinary stones treated by combined URS with holmium laser lithotripsy at a single institute were retrospectively evaluated. Correlations of possible predictors with SF status were analyzed using a logistic regression model. Two types of SA were measured: "Traced stone surface area'' (tSA) and "calculated stone surface area'' (cSA).
Results: According to the univariate analysis, the following variables were significantly associated with non-SF status: Stone number (P &lt; 0.001), ureteral stone location (P = 0.045), presence of renal stones (P &lt; 0.001), tSA (P &lt; 0.001), cSA (P &lt; 0.001), stone volume (P &lt; 0.001), and operator experience (P = 0.02). According to multivariate analysis, stone volume (P = 0.016) was an independent predictor of SF status. The scatter diagrams for tSA and cSA showed strong correlations between these parameters, and Spearman p was 0.975.
Conclusions: Stone volume and SA were highly indicative of stone status after single semirigid URS, with or without a flexible component. The formula for cSA, maximum diameter x width x p pi 1/4, was demonstrated to accurately represent SA in this study. SA, however, indicated a lower clinical priority and utility as a predictor of stone status than stone volume. The combination of semirigid and flexible URS could access any ureteral stones, including those that semirigid URS alone could not treat. The cutoff points for these predictors of outcome were 110.0mm(2) for cSA, 125.0mm(2) for tSA, and 840.0mm(3) for stone volume.

DOI： 10.1089/end.2012.0548

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Language：Japanese   Publisher：(一社)日本泌尿器科学会  

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Language：English  

Research on in vitro spermatogenesis has a long history and remained to be an unaccomplished task until very recently. In 2010, we succeeded in producing murine sperm from primitive spermatogonia using an organ culture method. The fertility of the sperm or haploid spermatids was demonstrated by microinsemination. This organ culture technique uses the classical air-liquid interphase method and is based on conditions extensively examined by Steinbergers in 1960s. Among adaptations in the new culture system, application of serum-free media was the most important. The system is very simple and easy to follow.

DOI： 10.1007/978-1-62703-038-0_41

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

Purpose: Azoospermia is a common side effect of chemotherapy. Although most patients restore spermatogenesis overtime, the exact time course has not been well described. We analyzed the recovery of spermatogenesis in testicular cancer patients following chemotherapy. Patients and Methods: 49 patients, consisting of 45 treated with a bleomycin, etoposide and cisplatin (BEP) regimen and 4 with high-dose chemotherapy, were followed up with occasional semen analyses. The primary endpoint of this study was the confirmation of motile spermatozoa in the patients' semen. Results: Among 45 patients treated with BEP, 44 recovered spermatogenesis. The recovery of spermatogenesis was delayed depending on the increase in BEP cycles. In groups of patients who received 1-2, 3 and 4 cycles, the recovery rates of spermatogenesis within 2 year were 83.3, 80.0 and 66.7%, respectively. In the group with 5-6 cycles of BEP, re-spermatogenesis was significantly delayed and no patients re-established spermatogenesis within 2 years. The patients' age and semen parameters before chemotherapy were not useful as predictive factors for the recovery of spermatogen-esis. Conclusion: The recovery of spermatogenesis was rather fast and was often observed as early as several months after BEP treatment when the number of cycles was &lt;4. Copyright (C) 2013 S. Karger AG, Basel

DOI： 10.1159/000351189

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Language：English   Publishing type：Research paper (scientific journal)  

Background: This study investigated the correlation between the operation time using two different power settings of a Ho: YAG laser. Findings. A total of 68 patients underwent cystolithotripsy from April 2010 to October 2011 In Fifty-six of these patients underwent cystolithotripsy by one surgeon using a Ho: YAG laser for bladder calculi. This study assessed these patients in two groups
the 30 W laser generator group with the settings of 2.5 J x 5 Hz (30 W group) and the 100 W laser generator group as the settings of 3.5 J x 5 Hz (100 W group). The operation time in these two groups were assessed.A total of 56 patients including 45 male and 11 female patients that underwent cystolithotripsy using a Ho: YAG laser for bladder calculi by one surgeon were enrolled in this study. The patients' characteristics including age (mean
68.8 vs 68.4 yr), gender (male
74.2 vs 88.0%), stone burden (mean
34.9 vs 41.3 mm), number of stones (mean
3.2 vs 2.0) and stone's CT density (mean
981.5 vs 902.0 HU) showed no significant differences. All patients were stone free following treatment. The median total length of the operation was 19 minutes (mean: 34.6 ± 36.1) in the 30 W group and 29 minutes (mean: 44.4 ± 38.8) in the 100 W group, which was not significantly different. Conclusions: The results showed that the power settings of Ho: YAG laser show no differences in the operation time for bladder calculi lithotripsy. © 2013 Kawahara et al
licensee BioMed Central Ltd.

DOI： 10.1186/1756-0500-6-80

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Language：English   Publisher：JAPANESE SOCIETY OF OVA RESEARCH  

It is a challenge to induce spermatogenesis <i>in vitro</i>, because it involves a complicated process of sequential cell proliferation and differentiation, from spermatogonial stem cells to sperm formation. Recently, we have succeeded, using a classical organ culture method, to produce sperm from spermatogonial stem cells, using several modifications. The produced spermatids and sperm were fertile, giving rise to healthy progeny. This new <i>in vitro</i> system for spermatogenesis could be useful for addressing the issue of sperm quality that is becoming more important in this ICSI era. We also hope that an <i>in vitro</i> system of human spermatogenesis will be developed in the near future.<br>

DOI： 10.1274/jmor.30.155

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Other Link： https://jlc.jst.go.jp/DN/JALC/10026590522?from=CiNii

DOI： 10.1371/journal.pone.0065060

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DOI： 10.1002/dvg.22391

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Other Link： http://orcid.org/0000-0002-6198-2234

DOI： 10.1091/mbc.E13-05-0234

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Language：English   Publisher：SPRINGER  

DOI： 10.1007/s00240-012-0486-y

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

The first indwelling ureteral splint was described in 1967. A ureteral stent can cause unpleasant side effects, such as urinary frequency, urgency, incontinence, hematuria, bladder pain and flank pain, which have a negative impact on a patient's quality of life. It is necessary to minimize the amount of material in the bladder in order to decrease stent-related symptoms. This study investigated the stent-related symptoms after changing from a double pigtail to a loop-type ureteral stent in the same patient group. This study followed 25 patients who underwent ureteral stent exchange from double pigtail to loop-type ureteral stent between September 2009 and February 2010. Ureteral stents were exchanged using topical, conscious sedation and general anesthesia for the various procedures including stent exchange, before/after shock wave lithotripsy and before/after ureteroscopy. The stent length was selected to be the same as whole ureteral length and the caliber based on the previous stent. A self-administered stent-related symptom questionnaire was used to assess stent-related symptoms in comparison to the previous double-pigtail stents. A total of 25 patients with a median age of 56.5 years underwent ureteral stent exchange. All patients had stone disease except two patients who had ureteral stricture. Almost all of stent-related symptoms without nocturia showed a significantly lower score with the loop-type ureteral stent than in double-pigtail stent. None of the patients experienced urinary tract infection either before or after undergoing ureteral stent exchange. Changing to loop-type ureteral stent significantly decreased ureteral stent-related symptoms.

DOI： 10.1007/s00240-012-0500-4

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE INC  

OBJECTIVE To evaluate the effectiveness of cystoscopic lithotripsy, we performed Amplatz sheath technique using Ho: YAG laser. Maheshwari first reported the use of an Amplatz sheath in the female urethra in 1998, and Okeke et al reported the use of an Amplatz sheath for male patients during cystolithotripsy in 2004. The usefulness of the holmium (Ho): yttrium aluminum garnet (YAG) laser lithotripsy is widely accepted, even for large bladder calculi. Since then, there have been no more reports of using the sheath with an Ho: YAG laser.
TECHNICAL CONSIDERATIONS We inserted the Amplatz sheath conversely. Because of the clear visualization, we used higher laser settings with 2.5 J x 15 to 20 Hz.
RESULTS We experienced 3 female patients that were successfully treated with the Amplatz sheath technique using Ho: YAG laser lithotripsy. In these 3 patients, whose stone burdens were 4.5, 3.8, and 4.3 cm, they were able to successfully become stone-free with surgeries of 74 minutes, 67 minutes, and 58 minutes, respectively, with no complications.
CONCLUSION We experienced 3 female patients that were successfully treated with the Amplatz sheath technique using Ho: YAG laser lithotripsy. UROLOGY 80: 1154-1155, 2012. (C) 2012 Elsevier Inc.

DOI： 10.1016/j.urology.2012.07.046

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATL ACAD SCIENCES  

Male infertility is most commonly caused by spermatogenic defects or insufficiencies, the majority of which are as yet cureless. Recently, we succeeded in cultivating mouse testicular tissues for producing fertile sperm from spermatogonial stem cells. Here, we show that one of the most severe types of spermatogenic defect mutant can be treated by the culture method without any genetic manipulations. The Sl/Sl(d) mouse is used as a model of such male infertility. The testis of the Sl/Sl(d) mouse has only primitive spermatogonia as germ cells, lacking any sign of spermatogenesis owing to mutations of the c-kit ligand (KITL) gene that cause the loss of membrane-bound-type KITL from the surface of Sertoli cells. To compensate for the deficit, we cultured testis tissues of Sl/Sl(d) mice with a medium containing recombinant KITL and found that it induced the differentiation of spermatogonia up to the end of meiosis. We further discovered that colony stimulating factor-1 (CSF-1) enhances the effect of KITL and promotes spermatogenesis up to the production of sperm. Microinsemination of haploid cells resulted in delivery of healthy offspring. This study demonstrated that spermatogenic impairments can be treated in vitro with the supplementation of certain factors or substances that are insufficient in the original testes.

DOI： 10.1073/pnas.1211845109

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARY ANN LIEBERT INC  

Purpose: To assess the utility of attenuation coefficients as predictors of surgical outcome after a single flexible ureteroscopy (URS) with holmium laser lithotripsy. Many reports indicate that the efficacy of extracorporeal shockwave lithotripsy (SWL) can be predicted by the target's radiofrequency attenuation, measured as Hounsfield units (HUs) on noncontrast CT (NCCT). Studies of flexible URS, however, have not assessed the predictive value of attenuation coefficients on NCCT.
Patients and Methods: Patients with renal stones who were treated by flexible URS with holmium laser lithotripsy between December 2009 and October 2011 at a single institute were retrospectively evaluated. Stone-free (SF) status was determined by kidneys-ureters-bladder (KUB) radiography at postoperative month 3. Correlations of possible predictors with SF status were analyzed using a logistic regression model. The comparison between groups with low and high HUs was examined using the Mann-Whitney U test.
Results: There were 219 eligible procedures. According to the logistic regression model, the maximum attenuation coefficient (P = 0.105) and average attenuation coefficient (P = 0.175) did not significantly, independently predict SF status. Fragmentation efficiency was significantly different between cases with low and high attenuation coefficients (P = 0.001). In groups with less than 20.0-mm diameter stones, overall operative time (P &lt; 0.001 and P = 0.001) and the time from starting fragmentation (P &lt; 0.001 and P = 0.002) were significantly high in both attenuation groups. In groups with stones greater than 20.0 mm diameter, the two definitions of operative time revealed no differences between the low and high attenuation groups. The retrospective study design was the major limitation of this study.
Conclusions: We found that both the maximum and average attenuation coefficients on NCCT are significantly related to the fragmentation efficiency. In addition, this study showed that, in patient groups with stone a burden &lt;20.0 mm in diameter, both the maximum and average attenuation coefficients were significantly predictive of operative time.

DOI： 10.1089/end.2012.0154

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Complete staghorn calculi are typically managed with percutaneous nephrolithotomy (PCNL). However, dilating nephrostomy and inserting a nephro access sheath can be difficult to perform without hydronephrosis. We reported the procedure of ureteroscopy-assisted retrograde nephrostomy (UARN) during PCNL. UARN is effective without dilating the renal collecting system in cases of complete staghorn calculi. A 63-year old female with a left complete staghorn renal calculus was referred to our hospital. Under general and epidural anesthesia, the patient was placed in a modified-Valdivia position. A flexible ureteroscope was inserted and a Lawson retrograde nephrostomy puncture wire was advanced into the flexible ureteroscope. The puncture wire was forwarded along the route from the renal pelvis to the exit skin. Calculus fragmentation was done using a pneumatic lithotripter and the Ho: YAG laser. UARN during PCNL was effective for the treatment of a complete staghorn calculus. Copyright © 2012 S. Karger AG, Basel.

DOI： 10.1159/000343519

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE INC  

OBJECTIVE To examine which parameters should be measured to preoperatively determine the stone burden as predictors of stone-free (SF) status after a single flexible ureteroscopy (URS). Although several stone parameters reportedly influence the outcome of treatment for urinary stones, the most reliable indicators of stone burden remain unclear.
METHODS Patients with renal stones treated by flexible URS with holmium laser lithotripsy between October 2009 and December 2011 at a single institute were retrospectively evaluated. The SF status was determined by kidney-ureter-bladder (KUB) films at postoperative day 1. Correlations of possible predictors with the SF status were analyzed using a logistic regression model.
RESULTS According to the univariate analysis, the following variables were significantly associated with failed treatment: number of stones (P = .001), cumulative stone diameter (CSD) (P &lt; .001), stone surface area (SA) (P &lt; .001), stone volume (P &lt; .001), and presence of lower pole calculi (P = .008). According to the multivariate analysis, the stone volume (P &lt; .001) and the CSD (P = .015) were found to be independent predictors of SF status. The SA (P = .598) had no significant independent influence on the SF status.
CONCLUSION Among the several parameters regarding the renal stone burden, the stone volume determined by noncontrast computed tomography and the CSD of the KUB were significantly and independently inversely related to the success rate of URS. Among the 3 parameters of stone burden, the SA was found to have a lower clinical utility and priority as a predictor of a SF status after URS. UROLOGY 80: 524-528, 2012. (c) 2012 Elsevier Inc.

DOI： 10.1016/j.urology.2012.04.001

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

A large renal stone can be treated ureteroscopically, but the treatment often requires more than one procedure. The use of stenting before ureteroscopy was recently reported. The present study investigated the effectiveness of preoperative stenting before ureteroscopic lithotripsy for large (&gt;15 mm) renal stones. A ureteral stent was intentionally inserted in 25 patients undergoing ureteroscopic surgery. A group of 36 non-prestented patients was used as control. Median stone diameter was 21 mm in both groups. Pre-ureteroscopy stenting significantly improved the stone-free rate, defined as stones &lt;2 mm and &lt;4 mm (P &lt; 0.05), whereas it did not significantly improve the stone-free rate defined as 0 mm (P = 0.12). The uretereoscopy success rate was 72.0% in the stented and 55.6% in the control group (P = 0.09). A 14/16-Fr ureteral access sheath was successfully inserted in 94.7% of the stented patients, and 74.2% of the non-stented patients (P &lt; 0.05). Our findings showed that preoperative stenting is effective for dilation of the ureter, and also to facilitate the insertion of a ureteral access sheath in patients undergoing ureteroscopic lithotripsy for large renal stones.

DOI： 10.1111/j.1442-2042.2012.03046.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

OBJECTIVE
To describe a technique for ureteroscopy assisted retrograde nephrostomy.
PATIENTS AND METHODS
Under general and epidural anaesthesia, the patient is placed in a modified-Valdivia position. Flexible ureteroscopy is carried out, and a Lawson retrograde nephrostomy puncture wire is placed in the ureteroscope (URS).
After the needle has exited through the skin, no further steps are required in preparation for dilatation.
RESULTS
After informed consent was obtained, two patients (a 43-year-old man with left renal stones and a 57-year-old woman with right renal stones) underwent this procedure.
The URS was positioned in the middle posterior calyx and punctured toward the skin.
CONCLUSIONS
This procedure involves less radiation exposure and shorter surgery than the previous percutaneous nephrostomy technique.
Our technique represents another new option for percutaneous nephrolithotomy in patients with a non-dilated intrarenal collecting system.

DOI： 10.1111/j.1464-410X.2011.10795.x

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Objective. This study investigated the correlation between the operation time and the stone size as determined by multiple modalities and the computed tomography (CT) densities of bladder calculi using holmium: yttrium garnet (Ho:YAG) laser lithotripsy. Material and methods. A total of 68 patients underwent cystolithotripsy from March 2010 to October 2011. Thirty-six of these patients underwent cystolithotripsy using a Ho:YAG laser for bladder calculi by a single surgeon. The stone size was assessed by six modalities: sum of the stones' diameters: stone burden; maximum stone's diameter; number of stones; sum of the area using axial CT; sum of area using kidney-ureter-bladder (KUB) films; and volume using CT. In addition, the stone's CT density was measured by: the mean CT density of the maximum stone's whole area; maximum CT density of the maximum stone's whole area; and mean CT density of the maximum stone's center area. Correlations between the operation time and the stone size and the stone CT density were assessed. Results. A total of 36 patients (30 male and six female) who underwent cystolithotripsy using a Ho:YAG laser for bladder calculi were enrolled in this study. Spearman correlation showed that the area and volume were strongly correlated with the operative time. The multipliers between the stone size and stone CT density showed no advantages based on the stone area or volume alone. Conclusion. The area and volume of the stones correlated more closely with the operation time than the stone burden for bladder calculi lithotripsy using a Ho:YAG laser.

DOI： 10.3109/00365599.2012.672456

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DOI： 10.1159/000342338

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Objectives: To define the best modality for estimating ureteral length in patients undergoing ureteral stent placement. Methods: This study enrolled 151 patients (169 ureters) undergoing stent insertion. In all of them, an intravenous urography and non-contrast computed tomography were carried out. The actual ureteral length was determined by direct measurement using a 5-Fr ureteral catheter. A multivariate analysis evaluated the association between the ureteral length and each of the following parameters: body height, body surface area, ureteral trace by intravenous urography, linear distance (liner distance 1) from the ureteropelvic junction to the ureterovesical junction by intravenous urography, linear distance (liner distance 2) from the mid kidney to the ureterovesical junction by intravenous urography, and the distance from the level of the renal vein to the ureterovesical junction by axial computed tomography (axial computed tomography distance). Results: The mean actual ureteral length was 23.2 cm (median 24 cm, range 1629 cm). The Spearman correlation coefficients for body height, body surface area, ureteral trace, liner distance 1, liner distance 2 and axial computed tomography distance were 0.3126, 0.3076, 0.4541, 0.5230, 0.4796 and 0.6168, respectively. Axial computed tomography distance showed the best correlation with the actual ureteral length. Conclusion: The axial computed tomography distance as calculated by the axial computed tomography can more reliably predict the actual ureteral length than other parameters. Further studies are required to show the best method for estimating the actual ureteral length in patients undergoing ureteral stent placement.

DOI： 10.1111/j.1442-2042.2012.02998.x

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DOI： 10.1111/j.1442-2042.2012.02984.x

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Language：English   Publisher：WILEY-BLACKWELL  

DOI： 10.1111/j.1442-2042.2012.02979.x

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Introduction: We developed a method for ureteral stent removal in female patients that requires no cystoscopy or fluoroscopic guidance using a crochet hook. In addition, we also investigated the success rate, complications and pain associated with this procedure.
Methods: A total of 40 female patients (56 stents) underwent the removal of ureteral stents. All procedures were carried out with the patients either under anesthesia, conscious sedation, or analgesic suppositories as deemed appropriate for each procedure including Shock Wave Lithotripsy (SWL), Ureteroscopy (URS), Percutaneous Nephrolithotomy (PCNL), and ureteral stent removal. At the time of these procedures, fluoroscopy and/or cystoscopy were prepared, but they were not used unless we failed to successfully remove the ureteral stent using the crochet hook. In addition, matched controls (comprising 50 stents) which were removed by standard ureteral stent removal using cystoscopy were used for comparison purposes.
Results: A total of 47 of the 56 stents (83.9%) were successfully removed. In addition, 47 of 52 (90.4%) were successfully removed except for two migrated stents and two heavily encrusted stents which could not be removed using cystoscopy. Ureteral stent removal using the crochet hook technique was unsuccessful in nine patients, including two encrustations and two migrations. Concerning pain, ureteral stent removal using the crochet hook technique showed a lower visual analogue pain scale (VAPS) score than for the standard technique using cystoscopy.
Conclusions: Ureteral stent removal using a crochet hook is considered to be easy, safe, and cost effective. This technique is also easy to learn and is therefore considered to be suitable for use on an outpatient basis.

DOI： 10.1371/journal.pone.0029292

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We encountered three patients with dysuria who had undergone spinal surgery for spina bifida during infancy. The patients presented with mental disability and dysbasia. Difficulty in urination, urinary incontinence, and a residual sensation of urine were resolved through intermittent self-catheterization in all patients. It was speculated that treatment for dysuria in these patients was delayed because they were not aware of its association with their condition during infancy, dysuria was relatively mild, and they had no history of febrile urinary tract infection. It is important for attending physicians to explain to parents of such infants the possible association of spina bifida with the future risk of dysuria, and to consider long-term follow-up to monitor their outcome.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

Purpose: The ureteral stent is now a fundamental part of many urological procedures. To decrease ureteral stent-related symptoms, loop type ureteral stent was developed. However the most important factor to decrease urinary symptoms is choosing the optimal length of a ureteral stent. We investigated the relationship between the actual ureteral length and the loop type ureteral stent position. Materials and Methods: A total of 226 loop type polyurethane ureteral stents (156 patients) were inserted with four options for stent length (20, 22, 24 and 26 cm). The ureteral length was measured using a ruled 5-Fr ureteral catheter. The appropriateness of stent position was defined into three groups based on kidney-ureter-bladder films. Results: Nine stents (3.9%) migrated, 171 stents (75.7%) were in the appropriate position and 46 stents (19.5%) were overlong. The rate of migration rate and overlong stents closely correlated with the ureteral length when the proximal end of the stent was in the renal pelvis. Conclusions: It is appropriate to choose a loop type ureteral stent that is the same or 1 cm less than the length of the ureter when the proximal end of the stent will be in the renal pelvis. Copyright (C) 2011 S. Karger AG, Basel

DOI： 10.1159/000332431

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Hindawi Limited  

A 23-year-old female had bilateral ureteral stents placed due to bilateral renal stones and hydronephrosis. The bilateral ureteral stents were changed every 3 months. A kidney ureter bladder (KUB) film showed left encrustation along the ureteral stent thus necessitating removal
however, the ureteral stent could not be removed cystoscopically. The ureteral stent was, therefore, extracted using flexible ureteroscopy (URS) with a holmium (Ho): yttrium aluminum garnet (YAG) laser. © 2012 Takashi Kawahara et al.

DOI： 10.1155/2012/862539

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Introduction. Open surgical anatrophic nephrolithotomy (ANL) had been the standard treatment for large renal calculi prior to the development of endoscopic devices and endoscopic techniques. A previous report described the efficacy of ureteroscopy-assisted retrograde nephrostomy (UARN) and presented a case of renal calculi successfully treated with UARN during percutaneous nephrolithotomy (PCNL) in a patient after ANL. Case Presentation. A 61-year-old male with left renal calculi was referred for further treatment. The patient was placed under general and epidural anesthesia, in a Galdakao-modified Valdivia position. A flexible ureteroscope (URS) was inserted, and a Lawson retrograde nephrostomy puncture wire was advanced into the flexible URS. The puncture wire then followed the route from the renal pelvis to the exit skin. Calculus fragmentation was undertaken using a pneumatic lithotripter. Conclusions. UARN for PCNL was therefore found to be a safe, effective, and appropriate treatment for a patient presenting with renal calculi after undergoing ANL. © 2012 Takashi Kawahara et al.

DOI： 10.1155/2012/164963

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Introduction: We previously reported on the effectiveness of ureteroscopy-assisted retrograde nephrostomy during percutaneous nephrolithotomy and report two cases of lower calyx calculi in horseshoe kidney that were successfully treated with ureteroscopy-assisted retrograde nephrostomy. During the ureteroscopy-assisted retrograde nephrostomy procedure, a ureteroscope is advanced in the desired calyx and a Lawson retrograde nephrostomy puncture wire is inserted. The wire is advanced through the calyx to exit the skin. The wire is then used for the percutaneous dilation. Case presentation: Case 1 was a 68-year-old man who was shown on radiography to have left lower calyx calculi (19 × 15mm, 7 × 5mm, and 7 × 3mm) in horseshoe kidney. Case 2 was a 36-year-old woman shown on radiography to have a left lower calyx calculus (10 × 8mm) in horseshoe kidney. Conclusions: Both patients were stone-free after ureteroscopy-assisted retrograde nephrostomy during percutaneous nephrolithotomy. Ureteroscopy-assisted retrograde nephrostomy is a promising procedure for safely and effectively treating lower calyx stones in horseshoe kidney. © 2012 Kawahara et al.
licensee BioMed Central Ltd.

DOI： 10.1186/1752-1947-6-194

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Purpose: We developed a method for ureteral stent exchange in female patients under fluoroscopic guidance using a crochet hook technique (CHEX). Patients and Methods: A total of 45 female patients (51 stents) underwent exchange of ureteral stents. In these patients, 21 ureteral stents were exchanged using CHEX. All procedures were carried out with the patients under conscious sedation. At the time of the procedures, we extracted the ureteral stent from the external urethral orifice using CHEX under fluoroscopic guidance and inserted the new stent under fluoroscopic guidance without cystoscopy. Results: 20 of the 21 stents (95.2%) were successfully exchanged. Ureteral stent exchange using CHEX was unsuccessful in 1 patient, including migration to the ureter. Conclusions: Ureteral stent exchange using a crochet hook is easy, safe and cost-effective. This technique was also easy to learn. Copyright (C) 2012 S. Karger AG, Basel

DOI： 10.1159/000336870

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Spermatogonial stem cells (SSCs) are the only stem cells in the body that transmit genetic information to the next generation. The long-term propagation of rodent SSCs is now possible in vitro, and their genetic modification is feasible. However, their differentiation into sperm is possible only under in vivo conditions. Here we show a new in vitro system that can induce full spermatogenesis from SSC lines or any isolated SSCs. The method depends on an organ culture system onto which SSCs are transplanted. The settled SSCs form colonies and differentiate up into sperm. The resultant haploid cells are fertile, and give rise to healthy offspring through micro-insemination. In addition, the system can induce spermatogenesis from SSCs that show spermatogenic failure due to a micro-environmental defect in their original testes. Thus, an in vitro system is established that can be used to correct or manipulate the micro-environmental conditions required for proper spermatogenesis from murine SSC lines.

DOI： 10.1038/ncomms1478

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Language：English   Publisher：SOC STUDY REPRODUCTION  

DOI： 10.1095/biolreprod.110.089342

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Spermatogenesis is one of the most complex and longest processes of sequential cell proliferation and differentiation in the body, taking more than a month from spermatogonial stem cells, through meiosis, to sperm formation(1,2). The whole process, therefore, has never been reproduced in vitro in mammals(3-5), nor in any other species with a very few exceptions in some particular types of fish(6,7). Here we show that neonatal mouse testes which contain only gonocytes or primitive spermatogonia as germ cells can produce spermatids and sperm in vitro with serum-free culture media. Spermatogenesis was maintained over 2 months in tissue fragments positioned at the gas-liquid interphase. The obtained spermatids and sperm resulted in healthy and reproductively competent offspring through microinsemination. In addition, neonatal testis tissues were cryopreserved and, after thawing, showed complete spermatogenesis in vitro. Our organ culture method could be applicable through further refinements to a variety of mammalian species, which will serve as a platform for future clinical application as well as mechanistic understanding of spermatogenesis.

DOI： 10.1038/nature09850

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.102.328_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.102.429_5

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It is now possible to make mouse spermatogonial stem cells (SSCs) proliferate in vitro. However, these cultured cells, called germ-line stem (GS) cells, consist of not only SSCs but also a greater number of progenitor spermatogonia. Moreover, isolated GS cells barely proliferate. To elucidate the nature of SSCs and progenitor spermatogonia, we adapted a microdrop culture system to GS cells. Using a micromanipulator, individual microdrops were seeded with clusters or dissociated known numbers of GS cells. The number of surviving colonies was determined after 30 days. The proliferation rate of GS cells in microdrops increased as the number of GS cells seeded increased. It was observed that as few as three GS cells seeded in a microdrop can proliferate and expand the colony size. Those GS cells of expanded colonies were able to proliferate following subculture and underwent spermatogenesis in the host testis after transplantation into the seminiferous tubules of recipient mice. These data revealed that SSCs can multiply in a microdrop culture system. Microdrop culture offers a novel tool to elucidate the nature of SSCs in regard to their self-renewing capacity and can serve as a monitoring system of culture conditions for the self-renewal of SSCs.

DOI： 10.1095/biolreprod.109.082800

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Language：Japanese   Publisher：医学図書出版(株)  

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Achieving mammalian spermatogenesis in vitro has a long history of research but remains elusive. The organ culture method has advantages over the cell culture method, because germ cells are in situ albeit the tissue as a whole is in vitro. The method was used in the 1960s and 1970s but encountered difficulties in inducing complete meiosis, i.e., in getting meiosis to proceed beyond the pachytene stage. In the present study, we reevaluated the organ culture method using two lines of transgenic mice, Acr-GFP and Gsg2 (haspin)-GFP mice, whose germ cells express green fluorescent protein (GFP) at the mid and end stages of meiosis onward, respectively. Immature testicular tissues from these mice, ranging from 4.5 to 14.5 days postpartum, were cultured on the surface of the medium, providing a liquid-gas interface. Culturing testicular tissues of all ages tested resulted in the expression of both Acr-and Gsg2-GFP. Round spermatids were identified by a combination of Gsg2-GFP expression, cell size, and the presence of a single nucleus with a dot stained by Hoechst. In addition, the chromosome number of one of such presumptive spermatids was found to be 20 by the premature chromosome condensation method. As our semiquantitative assay system using GFP expression grading was useful for monitoring the effects of different environmental factors, including temperature, oxygen concentration, and antiretinoic molecules, further improvement of the culture conditions should be possible in the future.

DOI： 10.1095/biolreprod.110.083899

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Language：English   Publisher：ELSEVIER SCIENCE INC  

DOI： 10.1016/j.urology.2010.02.034

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Language：Japanese   Publisher：医学図書出版(株)  

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A 82-year-old man was referred to our hospital for treatment of hormone-refractory prostate cancer with liver metastases. The obstruction of intrahepatic bile ducts due to the rapid growth of liver metastases induced liver dysfunction. We administered 25 mg/m2 docetaxel on dayl and 280 mg/body estramustine phosphate on day 1 to day 3, every 4 weeks. After two courses of this combined chemothrapy, the liver metastases were markedly reduced in size with the rapid improvement of liver function.

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A 73-year-old female was operated with radical nephrectomy and cholecystectomy for renal cell carcinoma and suspected gallstones after 9 courses of sunitinib treatment. Gallbladder specimen showed gallbladder metastasis originating from the renal cell carcinoma. Gallbladder metastasis from renal cell carcinoma is rare. Here, we discuss a case of gallbladder metastasis from renal cell carcinoma. Copyright © 2010 S. Karger AG.

DOI： 10.1159/000279308

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.101.484_3

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The case was a 67-year-old male who visited our hospital with a major complaint of macroscopic hematuria. A bladder tumor was found. When a transurethral resection of the bladder tumor was performed, the histopathological diagnosis was neuroendocrine bladder cancer. After chemotherapy with cisplatin and etoposide a partial shrinkage of the tumor was observed
however, the patient expired 7 months after the first visit. Copyright © 2010 S. Karger AG.

DOI： 10.1159/000289584

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.101.519_2

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DOI： 10.5980/jpnjurol.101.430_3

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DOI： 10.1159/000245926

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A 54-year-old woman consulted our hospital for further evaluation of an incidentally detected small renal mass. Abdominal computed tomography (CT) and other imaging showed a right renal mass adjacent to the Riedel's lobe. Laparoscopic right partial nephrectomy was performed under a diagnosis of suspected renal cell carcinoma. Riedel's lobe is an anatomic variant, which is a caudal extension of the right lobe of the liver. Riedel's lobe presented an obstruction during right renal surgery, but we performed the procedure successfully because we retracted the lobe medially. This procedure yielded a clear surgical field, allowing us to perform this delicate operation safely.

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Mixed epithelial and stromal tumor of kidney (MEST-K) is a rare benign renal tumor that was first described by Michal and Syrucek in 1998. Its frequency is 0.2-0.28% of all the renal tumors. Here, we report an additional case of MEST-K occurring in a 28-year-old woman. The patient visited a hospital with complaints of lumbago and fever caused by pyelonephritis. The computed tomography revealed hydronephrosis and a cystic tumor in the right kidney, and laparoscopic right nephrectomy was performed. The resected kidney contained a cystic lesion with a grayish-white mural nodule, in the lower portion. The entire lesion measured 5 cm in diameter, and the mural nodule 2.5 cm in diameter. Histologically, the cyst was lined with tall columnar and transitional epithelia. The mural nodule showed microcystic architectures lined with tall columnar and transitional epithelia, scattered in a compact stroma. Immunohistochemically, spindle cells in the stroma were positive for smooth muscle-specific actin, and estrogen and progesterone receptors (ER and PR). Based on these findings, the tumor was diagnosed as MEST-K. MEST-K was newly introduced to the WHO classification of renal tumors, with a pathogenesis related to long-term estrogen exposure, because of ER and PR expression in the stroma. It is important to consider the possibility of this tumor when encountering cases of cystic tumor in middle-aged and older women, and men with a previous history of estrogen administration.

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18F-FDG PETによる前立腺がんの診断は、従来難しいと考えられてきた。CTとの融合によるPET-CTの導入で、前立腺がんの診断向上の可能性があると考えられ前立腺がんの診断の有用性について検討した。PSA高値(PSA平均値17.7ng/ml)の患者27名に対して、前立腺針生検の前にPET-CTを施行し、病理結果との比較検討を行った。前立腺がんが見つかった患者でのPET-CTの陽性率は37%であった。臨床病期別でPET-CTでの陽性率は、T1cでは25%、T2aでは50%、T2cでは67%、T3aでは100%であった。T2以上の比較的進行した前立腺がんでのPET陽性率は高く、有用性が高いと考えられた。(著者抄録)

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A 28-year-old man with adult-onset idiopathic male hypogonadotropic hypogonadism (MHH) is reported. He had been delivered normally and had normal puberty. He was referred to our hospital with a chief complaint of infertility. Serum levels of testosterone, luteinizing hormone, and follicle stimulating hormone (FSH) were low. Semen analysis demonstrated azoospermia. Pituitary hypofunction was suggested by gonadotropin releasing hormone (GnRH) loading test. Magnetic resonance images did not detect any abnormalities in the hypothalamic-pituitary region. After a diagnosis of adult-onset hypogonadotropic hypogonadism was established, the patient received human chorionic gonadotropin (hCG) and recombinant FSH treatment. After 5 months, his sperm count reached 6.9 X 10 6 per ml and his wife became pregnant. Adult-onset HH in most cases is caused by tumors and trauma. To our knowledge 17 cases of adult-onset idiopathic HH have been reported, and there were only 3 cases that were caused by pituitary dysfunction. This report showed that r-FSH and hCG therapy was effective in promoting fertility in a patient with adult-onset idiopathic MHH.

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We report a case of colorectal cancer with metastasis to the upper urinary tract. A 56-year-old man had left flank pain. Ultrasonography and computed tomographic (CT) examination demonstrated left hydronephroureter and a soft-tissue structure within the left ureter. Urinary cytology of the left ureter showed class IIIb. We diagnosed him with ureteral cancer and performed left nephroureterectomy. Microscopic examination demonstrated adenocarcinoma located in ureteral and pelvic wall, especially in blood vessels, with intact mucosa and similar to adenocarcinoma of colon cancer. Therefore metastatic upper urinary tract tumor was suspected. Barium enema and positron emission tomography-CT demonstrated sigmoid colon cancer. Biopsy specimen of colon cancer demonstrated adenocarcinoma, which was consistent with the ureteral tumor. Finally we diagnosed him with metastatic upper urinary tract tumor of sigmoid colon cancer. (Hinyokika Kiyo 55 : 339-343, 2009).

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A 66-year-old woman, who developed pulmonary tuberculosis at 17 years old, had a high fever in December, 2006. Computed tomographic (CT) scan showed a tumor in the left chalked kidney, which measured 7 cm in diameter with very low enhancement. Laboratory data showed the rise of acute phase reactants (erythrocyte sedimentation rate and c-reactive protein) and severe anemia. The cultures of sputum and urine revealed no Mycobacterium tuberculosis. With the diagnosis of left renal cell carcinoma in the chalked kidney, we performed left radical nephrectomy. Histopathological diagnosis was sarcomatoid renal cell carcinoma. Although sarcomatoid renal cell carcinoma is highly malignant and its prognosis is poor, her post-operative condition has been good without any adjuvant treatments and there have been no recurrent or metastatic lesions for 9 months. The supervention of renal cell carcinoma on renal tuberculosis is rare. The possible effects of tuberculous lesions on the development and progression of renal cell carcinoma are discussed. (Hinyokika Kiyo 55 : 253-257, 2009).

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Fragments of testis tissue from immature animals grow and develop spermatogenesis when grafted onto subcutaneous areas of immunodeficient mice. The same results are obtained when dissociated cells from immature testes of rodents are injected into the subcutis of nude mice. Those cells reconstitute seminiferous tubules and facilitate spermatogenesis. We compared these two methods, tissue grafting and cell-injection methods, in terms of the efficiency of spermatogenesis in the backs of three strains of immunodeficient mice, using neonatal porcine testicular tissues and cells as donor material. Nude, severe combined immunodeficient (SCID) and NOD/Sh-iSCID, IL-2R gamma(null)(c) (NOG) mice were used as recipients. At 10 months after surgery, the transplants were examined histologically. Both grafting and cell-injection methods resulted in porcine spermatogenesis on the backs of recipient mice; the percentage of spermatids present in the transplants was 67% and 22%, respectively. Using the grafting method, all three strains of mice supported the same extent of spermatogenesis. As for the cell-injection method, although SCID mice were the best host for supporting reconstitution and spermatogenesis, any difference from the other strains was not significant. As NOG mice did not show any better results, the severity of immunodeficiency seemed to be irrelevant for supporting xeno-ectopic spermatogenesis. Our results confirmed that tubular reconstitution is applicable to porcine testicular cells. This method as well as the grafting method would be useful for studying spermatogenesis in different kinds of animals.

DOI： 10.1038/aja.2008.5

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

We intended retrospectively to investigate whether reactive oxygen species (ROS) levels, detected in whole semen, were correlated with the actual pregnancy rate.
A total of 89 patients with data of ROS in semen, attending our male infertility clinics from April 1994 to June 2000, were evaluated. Semen parameters were determined with computer assisted semen analyzer (CASA) and ROS production levels were measured using a computer-driven LKB Wallac 1251 Luminometer after the addition of 40 mu L of 4 mM luminol at the patients&apos; first visits. All of the participants were inquired about their partners&apos; pregnancies after the mean follow-up of 24.0 months (range 1.4 to 74.1). They were divided into two groups (pregnant group: n = 41, non-pregnant group: n = 48) and their characteristics, semen profiles and integrated ROS levels were analyzed.
There was no difference between the pregnancy rate of ROS detectable cases and negative cases. However, the mean integrated ROS level in detectable cases of the non-pregnant group was significantly higher than that in detectable cases of the pregnant group (115.61 +/- 74.32 mV/30 min/10(6) sperm versus 7.22 +/- 4.69 mV/30 min/10(6) sperm, P = 0.0033). Then, by calculating the receiver operating characteristics curve with 95% confidence intervals, 4.35 mV/30 min/10(6) sperm was considered as a cut-off value of ROS in semen for pregnancy.
These results indicate that (i) highly detectable ROS in whole semen of infertile patients may have implications in their partners&apos; pregnancies and that (ii) detection of ROS in whole semen has a prognostic value for idiopathic male infertility.

DOI： 10.1111/j.1442-2042.2008.02213.x

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.318_2

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.430_3

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.467_1

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DOI： 10.5980/jpnjurol.100.278_4

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Language：Japanese   Publisher：JAPANESE SOCIETY OF OVA RESEARCH  

Mammalian spermatogenesis is a classic adult stem cell system. Identification of a crucial self renewal factor, glial cell line derived neurotrophic factor (GDNF), has provided new opportunities in cell culture systems of testicular cells. Spermatogonia can be maintained for years in culture with GDNF. Transplantation experiments of cultured spermatogonia have shown that they are able to reconstitute all the germ cells with proper programming in the testes, suggesting that the stem cell population of spermatogonia is maintained <i>in vitro</i> for at least, a couple years. Recently, it has also been reported that cultured spermatogonia have the potency to convert to multi-potent stem cells. The rapid progress of <i>in vitro</i> culture systems of spermatogonia makes it possible to apply the culture systems not only to the study of male infertility but also to regenerative medicine.<br>

DOI： 10.1274/jmor.26.178

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.354_2

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

Background: In the present study, we compared 12-with 8-core biopsy in patients with prostate-specific antigen (PSA) levels of 4.0-20.0 ng/ml. We also examined whether the free/total (F/T) PSA ratio is useful for cancer detection in 12-core biopsy. Methods: A total of 419 men with PSA level between 4.0 and 20.0 ng/ml underwent transrectal ultrasound-guided transperineal needle biopsies of the prostate. Of these men, 235 underwent 8-core biopsy and 184 underwent 12-core biopsy. We compared the cancer detection rate between the 8- and 12-core biopsy groups by analyzing the PSA value, and especially the F/T PSA ratio. Results: The cancer detection rate in the 12-core group (35.9%) was significantly higher than in the 8-core group (23.8%). In cases of PSA level of 4.0-20.0 ng/ml with F/T PSA ratio less than 0.11, the cancer detection rate was 53.1% in the 12-core biopsy group. Performing 12-core biopsy resulted in a marked difference of cancer detection rate between men with F/T PSA ratio less than 0.11 and those with more than 0.12 in gray zone PSA (48.2% and 17.5%, respectively). Conclusions: Twelve-core biopsy can achieve a higher detection rate of prostate cancer than 8-core biopsy using F/T PSA ratio. Copyright (C) 2009 S. Karger AG, Basel

DOI： 10.1159/000209358

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.475_4

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.431_4

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.242_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.100.424_1

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Language：Japanese   Publishing type：Research paper (scientific journal)  

Silodosin (URIEF®), a new so-called 3rd generation alpha-1 blocker, is widely expected to be effective and useful for lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH), due to its high specificity to alpha-1 A receptor. We evaluated the efficacy of Silodosin, on 187 males 50 years old or over with the diagnosis of BPH. Silodosin significantly improved the International Prostate Symptom Score (IPSS) and quality of life (QOL) score from the day after administration was started. Among 166 patients whose data were available for the analysis of efficacy of Silodosin, 77.5% showed apparent subjective improvement. Eighty three patients, who had been taking another alpha-1 blocker but without satisfactory effects, showed almost the same improvements in IPSS and QOL score after switching to Silodosin as the remaining 83 patients who had no preceding treatment with an alpha-1 blocker. The improvements were not only in voiding symptoms but also in storage symptoms. The patients, who had serious storage symptoms, responded rather well to Silodosin and showed significant improvement. Taken together, Silodosin showed a quick effect for improving subjective symptoms and QOL, and was found to be useful for the management of LUTS with BPH.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

Embryonic germ-line cells are unipotent cells that give rise to either sperm or oocytes. However, pluripotent stem cells can be derived from primordial germ cells (PGCs) or spermatogonia, suggesting that germ-line cells retain a capacity for pluripotency. Here, we made genome-wide comparisons of the gene expression profiles of freshly isolated PGCs, in vitro-formed PGCs (iPGCs), and other stem cell lines, including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and germ-line stem (GS) cells. Comparing PGC with ESC, 382 genes/transcripts were significantly up-regulated in ESC, while 188 were elevated in PGC. This suggests that PGCs possess transcription program distinct from that of ESC, although both share expression of many pluripotency-associated genes.
Our micro-array analysis showed that the analyzed samples could be classified into two groups: one consisting of all the ESCs and most of EGCs, and the other containing PGC samples, iPGC, one type of female EGC and GS cells. We then identified "signature" genes for the two groups, and used them to characterize GS cells, EGC, and iPGCs, and revealed developmental status of each cell type. The relationships between PGCs and stem cells derived from embryos or germ cells are discussed in light of these findings.

DOI： 10.1111/j.1365-2443.2008.01211.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARY ANN LIEBERT INC  

Germ-line stem cells (GSCs) constitute a stem cell population with remarkable stability and proliferative potential in vitro and are a useful model for studying the mechanism of self-renewal and "stemness" function of committed tissue stem cells. To identify GSC-specific genes, we performed subtractive hybridization using cDNA from GSCs, testis, and embryonic stem (ES) cells, and successfully identified 11 genes highly expressed in GSCs. Histological analysis confirmed expression of Cry alpha b, Mcpt8, Cxcl5, Fth1, Ctla2 alpha, and Spp1 in undifferentiated spermatogonia on the basement membrane area of the seminiferous epithelium of the testis, where the GSC niche is thought to be located. Among GSC-specific genes, quantitative PCR analysis showed seven genes-Fth1, Cryab, Spp1, Bcap31, Arhgap1, Ctla2a, and Serpina3g-to be common transcripts highly expressed in hematopoietic stem cells (HSCs). Histological analysis confirmed that Ctla2 alpha-, Serpina3g-, and Spp1-expressing cells were observed in the trabecular bone region of the bone marrow, where the HSC niche is located. Furthermore, histological analysis revealed that only Spp1 was expressed in the hair follicle bulge in the area of the hair follicle stem cell niche. Thus, identifying stemness genes by comparative analysis to GSCs is a powerful tool with which to explore the fundamental commonalities of HSCs and other stem cell types.

DOI： 10.1089/scd.2007.0077

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Language：English   Publisher：BLACKWELL PUBLISHING  

Germ cells are defined by their innate potential to transmit genetic information to the next generation through fertilization. Males produce numerous sperm for long periods to maximize chances of fertilization. Key to the continuous production of large numbers of sperm are germline stem cells and their immediate daughter cells, functioning as transit amplifying cells. Recently, it has become possible to expand germline stem cells of rodents in vitro. In addition, multipotent stem cells, which are functionally the same as embryonic stem cells, have been established from neonatal mouse testes. These stem cells derived from the testis should contribute to biological research and technologies. On the other hand, the nature of human spermatogenesis is largely unknown due to the lack of an appropriate experimental system. However, the prevailing testicular sperm extraction procedure unraveled hitherto unknown facets of human spermatogenesis. The establishment of a culturing method for human spermatogonial stem cells in hopefully the near future would be a great benefit for achieving further insight into human spermatogenesis and should lead to more sophisticated diagnostic and therapeutic clinical measures for male infertility.

DOI： 10.1111/j.1442-2042.2007.01963.x

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Language：Japanese   Publisher：(公社)神奈川県医師会  

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.99.343_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.99.400_3

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Other Link： http://search.jamas.or.jp/link/ui/2007180306

Language：Japanese   Publishing type：Research paper (scientific journal)  

A chromosomal survey using the G-banding technique was performed on 87 subfertile male whose semen analysis demonstrated severe oligospermia and azoospermia at Yokohama City University Hospital between January 1990 and October 2002. Fourteen of these subjects demonstrated major chromosomal anomalies (16.1%). Semen analysis in these cases demonstrated azoospermia, except in one case of autosomal abnormality. Twelve patients showed sex chromosomal abnormalities including 8 Klinefelter syndrome (47XXY) and 2 XX males (46XX) and two patients had autosomal abnormalities. The follicle-stimulating hormone (FSH) value in these patients, except for the two cases of autosomal abnormality and one case of 46XYq-, was much higher than normal. Histological examination was performed in 7 cases. In these cases, intratesticular spermatozoa were seen in only two cases (Klinefelter syndrome case and ring chromosome 21 case). Chromosome studies are important in the evaluation of subfertile patients with azoospermia and severe oligospermia. Because the abnormal genotype could be transferred to the next generation, the importance of chromosome studies before ICSI should be emphasized.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

Testicular germ cell transplantation into the seminiferous tubules is at present the only way to induce spermatogenesis from a given source of spermatogonial stem cells. Here we show an alternative method that harnesses the self-organizing ability of testicular somatic cells. The testicular cells of embryonic or neonatal mice or rats and of newborn pigs were dissociated into single cells. Each of them reorganized into a tubular structure following implantation into the subcutis of immunodeficient mice. When mouse germline stem (GS) cells derived from spermatogonial stem cells and expanded in culture were intermingled with testicular cells of rodents, they were integrated in the reconstituted tubules and differentiated beyond meiosis into spermatids. Normal offspring were produced by the microinjection of those spermatids into oocytes. This method could be applicable to various mammalian species and useful for producing functional gametes from GS cells in a xenoectopic environment.

DOI： 10.1095/biolreprod.106.056895

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Language：Japanese   Publishing type：Research paper (scientific journal)  

We report a rare case of uterine corpus metastasis from superficial bladder cancer. A 78-year-old female presented with abnormal vaginal bleeding. She received transurethral resection of bladder tumor (TUR-Bt) two years previously, and the pathological findings were transitional cell carcinoma (TCC) grade 3 pT1. Eight courses of BCG instillation were performed postoperatively. There was no recurrence of bladder cancer when vaginal bleeding occurred. Cytology of vaginal discharge was class V, and transitional cell carcinoma suspected. Pathological finding of transvaginal uterine corpus biopsy was TCC. We diagnosed metastases to uterine corpus from bladder cancer.

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.98.420_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.98.300_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.98.411_4

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Japanese Urological Association  

We reported a case of laparoscopic heminephroureterectomy for ureter cancer in a horseshoe kidney. A 59-year-old woman presented with frequency and diagnosed as left lower ureter cancer with a horseshoekidney. We performed transperitoneal laparoscopic nephroureterectomy. Feeding vessels were four arteries and two veins. Isthmus of the horseshoe kidney was divided using LCS and hemostasis was made using monopolar shears. Operating time was 300 minutes. Total Blood loss was 400 ml. Laparoscopic pyeloplasty or ishmusectomy to benign disease of the horseshoe kidney is often reported, but that of heminephrectomy to malignancy is seldom reported. Laparoscopic Heminephectomy for a horseshoe kidney is difficult surgery for aberrant vessels and isthmus, so it tends to be avoided for safety. But if anatomical consideration about aberrant vessels etc is well done and we operate carfully, so we will be able to do it for safety and small invasive opration.

DOI： 10.5980/jpnjurol1989.98.786

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Language：Japanese   Publishing type：Research paper (scientific journal)  

We investigated the feasibility, safety, and anti-tumor activity of combination chemotherapy with gemcitabine and nedaplatin (GN therapy) in patients with urological cancer. A total of 12 patients were enrolled in the current study. Histological characteristics in this study were 9 patients of urothelial carcinoma (UC), 2 patients of squamous cell carcinoma of the ureter (SCC), and 1 patient of renal collecting-duct carcinoma (CDC). All UC and one SCC were treated with GN therapy as second line chemotherapy. Other 2 patients were treated as induction chemotherapy. Systematiccombination chemotherapy consisting of gemcitabine (1,000 mg/m 2 on day 1 and 8) and nedaplatin (80 mg/ m 2 on day 1) was performed. 5 partial responses and 7 stable diseases were achieved in 12 assessable patients and corresponded to an over all response rate of 42%. 2 (22%) of 9 patients of UC, all patients of SCC and CDC responded to GN therapy. Grade 3 and 4 toxicity was primarily hematologic. The result of current study suggests that GN therapy may be an effective treatment for patients with SCC and CDC. On the other hand, UC as second line chemotherapy was notsuccessful.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：BLACKWELL PUBLISHING  

Aim: It has been proposed that gonadotropin-releasing hormone (GnRH) analog administered after testicular damage stimulates the recovery of spermatogenesis. However, GnRH analogs suppress the function of sex accessory organs. In this study, we investigated whether testosterone also stimulates the regeneration of rat spermatogenesis after exposure to busulfan.
Methods: Male Fisher rats were divided into three groups of five each and all rats were treated with busulfan, 25 mg/kg, intraperitoneally at week 0. Group A served as the control. The other two groups received testosterone enanthate, 8 mg/kg, subcutaneous injections at 3 week intervals two times before (group B) or three times after (group C) busulfan. States of spermatogenesis were evaluated by histology and by the number of spermatid nuclei per testis at week 25.
Results: The mean percentage of 'recovered' seminiferous tubules plus or minus standard deviation was 10.3 +/- 7.8% in group A and 2.1 +/- 1.2% in group B. In both groups, more than 80% of the tubules remained degenerated. However, testes of group C rats showed an improvement of up to 37.1 +/- 20.5% (P &lt; 0.05). The significant recovery of spermatogenesis was also demonstrated in group C by counting the number of spermatid nuclei per testis ([78.8 +/- 57.5] x10(6)). However, the count was only (7.6 +/- 13.5) x10(6) and (0.52 +/- 1.0) x10(6) in group A and B, respectively.
Conclusions: Testosterone administration after severe testicular damage enhanced the regeneration of spermatogenesis in rats. We assumed that supplementary doses of testosterone would be more practical for clinical application than GnRH analogs, because exogenous testosterone maintains androgenicity.

DOI： 10.1111/j.1442-2042.2006.01484.x

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Language：Japanese   Publishing type：Research paper (scientific journal)  

In August 2000, a 62-year-old woman presented to another municipal hospital with macroscopic hematuria. Transurethral resection of bladder tumor (TUR-Bt) was performed. The pathological diagnosis was transitional cell carcinoma (TCC), G2 &gt
squamous cell carcinoma (SCC). TUR-Bt repeated in July 2003 indicated recurrence. The pathological diagnosis was TCC, G2. She was referred to our hospital in August 2003 because she desired bladder preservation. After cystoscopy and random biopsy, pathological diagnosis was TCC with squamous differentiation, G1-G2, pTis. She received 7 weekly intravesical bacillus Calmette-Guerin (BCG) instillations. In April 2004, TUR-Bt was repeated and multiple recurrences were found. The pathological diagnosis was TCC with squamous differentiation, G1-G2, pTa. She received 10 weekly intravesical Pirarubicin hydrochroride instillations. In August cystoscopy and random biopsy were performed for evaluation of the intavesical instillation treatment. Pathological diagnosis was atypical squamous cells. In November, cystoscopy revealed recurrence of a bladder tumor. After admission, a small papillary tumor and multiple flat lesion biopsies demonstrated SCC without obvious invasion. The patient underwent cystectomy. There were widespread areas of full thickness squamous atypia. Most of the bladder did not show appearance of typical TCC, but the final pathological diagnosis was TCC because the case developed from TCC and could not be diagnosed as pure SCC. The diagnosis of SCC in situ of bladder is difficult, and this may contribute to its rarity.

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Language：Japanese   Publisher：神奈川県医師会  

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Language：Japanese   Publishing type：Research paper (scientific journal)  

To investigate how urinary frequency and incontinence affect the patient's subjective quality of life (QOL) and whether an improvement in objective findings by medical treatment affects his/her subjective QOL, a voiding diary using the King's Health Questionnaire (KHQ) and International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF) was delivered to patients with urinary frequency and/or incontinence before and after treatment with propiverine hydrochloride for 8 weeks. Sixty-eight patients completed the diary and the questionnaires. Objective symptoms decreased significantly with respect to the mean frequency of urination and to the mean incidence of urinary incontinence. The KHQ and ICIQ-SF scores improved significantly with respect to all domains except personal relationships in the KHQ. In the KHQ, furthermore, a significant correlation was found between decreased incidence of urinary incontinence and improvement in role limitations and between decreased incidence of urinary incontinence and improvement in emotional problems. In the ICIQ-SF, a significant correlation was found between decreased incidence of urinary frequency and subjective improvement in quantity of leakage, between decreased incidence of urinary frequency and improvement in subjective QOL scores, between decreased incidence of urinary frequency and improvement in the total ICIQ-SF score, and between decreased incidence of urinary incontinence and improvement in subjective QOL scores. Thirty-two episodes of adverse reactions were observed. None of them were serious. These results suggest that an improvement in objective symptoms with propiverine hydrochloride favorably improves subjective QOL of the patient, and provide further evidence about the safety and efficacy of propiverine hydrochloride.

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A 46-year-old man complained of lower abdominal pain, and his abdominal and pelvic computed tomographic scan revealed left hydronephrosis and a huge tumor (9 × 9 cm) in the left distal ureter involving the left iliac vessel that was considered unresectable. Histological diagnosis showed squamous cell carcinoma, and histoculture drug response assay (HDRA) suggested the effectiveness of gemcitabine and nedaplatin. A cycle of adjuvant chemotherapy consisting of MEG (methotrexate 30 mg/m 2: day 1 and 15, epirubicin 50 mg/m 2: day 1, and cisplatin 50 mg/m 2: day 2 and 3) was performed as a first line chemotherapy, but the size of the ureteral tumor did not change. He was treated with 3 cycles of systematic combination chemotherapy consisting of gemcitabine (1,000 mg/m 2: day 1 and 8) and nedaplatin (80 mg/m 2: day 1). After 2 courses of chemotherapy, the tumor size was reduced by 50% (PR
RECIST guidelines) and the tumor markers (SCC, CYFRA, NSE, CEA, and CA19-9) dropped to within the normal range. There were no serious adverse events except for grade 3 neutropenia which spontaneously recovered. However, because the tumor size was not reduced after the third cycle of chemotherapy, we applied external beam radiation to the primary lesion and the metastatic retroperitoneal lymph node site. No evidence of residual tumor progression has been found for 6 months after radiation therapy. We concluded that GN chemotherapy may be useful for patients with squamous cell carcinoma of the ureter.

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.97.543_1

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER TOKYO  

The theory of the &apos;&apos; stem cell niche &apos;&apos; was originally proposed for the hematopoietic system, and the existence of the niche as an actual entity was proved in the Drosophila germ cell system. Historically, mammalian spermatogenesis has been studied extensively as a prime example of a stem cell system, and studies have established a stem-progenitor hierarchical order of spermatogonia. In the niche on the basal lamina of seminiferous tubules, spermatogonial stem cells (SSCs) are secluded from the outside world and divide constantly to self-renew and differentiate. During the last 10 years, the development and exploitation of the germ cell transplantation method has expanded our understanding of the nature of SSCs and their niches. The ability to maintain and expand SSCs in vitro, which recently became possible, has further reinforced this research area as a mecca of stem cell biology Nonetheless, the mammalian germ stem cell and its niche remain to be defined more strictly and precisely. We are still on a journey in search of the real stem cell and its true niche.

DOI： 10.1532/IJH97.05088

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BACKGROUND: Although patients with prostate cancer with metastatic lesions initially respond to androgen ablation therapy, most patients ultimately develop a hormone-refractory state. Effective treatment for men with hormone-refractory prostate cancer (HRPC) has not been established. We performed a clinical study of docetaxel in HRPC patients, and evaluated its efficacy. METHODS: Nine patients with HRPC were administered 55 mg/m2 docetaxel, every 3 weeks, simultaneously with hormonal therapy, with a luteinizing hormone-releasing hormone analog, and daily oral dexamethasone. Change in serum prostate-specific antigen (PSA) was determined as the primary endpoint. RESULTS: The mean age of the patients was 64 years (range, 49 to 76 years). Median follow-up time was 8.5 months (range, 5.3 to 16.7 months). In eight patients whose pretreatment serum PSA was elevated, six patients (75.0%) had a PSA decline of more than 50%, and four (50.0%) had a PSA decline of more than 75%. Median time to progression for all patients was 7.9 months (range, 0.0 to 11.6 months; 95% confidence interval [CI], 0.0 to 26.3). The median overall survival was 8.5 months (range, 5.3 to 16.7 months; 95% CI, 8.1 to 13.8). Four of six patients (66.7%) with pain before treatment obtained pain relief and were able to discontinue analgesic agents. This regimen was well tolerated. Grade 3 or 4 neutropenia or leukocytopenia without fever was seen in three patients (33.3%). Only one patient required administration of granulocyte-colony stimulating factor because of neutropenia. No other grade 3 or 4 toxicity was observed. CONCLUSION: Docetaxel was an active agent in Japanese HRPC patients, and was well tolerated in this population. To establish its efficacy and safety in Japanese HRPC patients, a large-scale study in Japan is warranted.

DOI： 10.1007/s10147-005-0490-0

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DOI： 10.1111/j.1440-1827.2005.01837.x

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DOI： 10.1111/j.1440-1827.2005.01813.x

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Language：Japanese   Publishing type：Research paper (scientific journal)  

Two patients with testicular tumors whose serum α-fetoprotein (AFP) persisted to show an abnormally high concentration are reported. Case 1 : A 42-year-old male who had been suffering from chronic hepatitis, underwent left high orchiectomy for a left testicular tumor in 1998. Diagnosis was an authentic stage I seminoma. In 2002, chemotherapy was performed for a metastatic seminoma revealed as a solitary mass in the mediastinum by radiographic studies, and histologically confirmed to be a metastatic seminoma. Although lymph nodes were gradually reduced in size, the serum AFP and transaminase levels remained at an abnormally high concentration. The subfraction profile with lens culinaris hemagglutinin (LCA) revealed elevation of only peak 1 which implied that the chronic hepatitis was due to liver dysfunction. After a 10-month follow-up the levels of both AFP and transaminase decreased, and the patient was disease-free. Case 2: In 2002, a 30-year-old male underwent left high orchiectomy for a left testicular tumor, and histological examination revealed seminoma, immature and mature teratoma, embryonal carcinoma. The serum AFP was elevated to 45 ng/ml. Diagnosis was authentic stage I. After 2 courses of chemotherapy, the serum AFP remained at an abnormally high concentration. However, there were no new lesions. The serum AFP level was not elevated in any of the family members. The subfraction profile with LCA revealed elevation of only peak 1, which implied that there were no viable lesions. After a 24-month follow-up AFP was about 20 ng/ml and the patient was disease-free.

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.96.133_3

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.96.242_1

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DOI： 10.5980/jpnjurol.96.184_2

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.96.142_4

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.96.285_3

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：INT SOC HISTOLOGY & CYTOLOGY  

Cnot7 is a co-factor of transcription regulation, expressed in a variety of tissues including the lung, liver, thyroid gland, and testis. Our previous study (Nakamura et al., 2004) showed that deletion of the Cnot7 gene in mice caused almost no abnormal phenotypes except for male infertility, due to oligo-astheno-teratozoospermia. This study also showed that Cnot7(-/-) mouse germ cells transplanted as donors could colonize in recipient wild mouse testes to develop normal spermatogenesis by spermatogonial transplantation assay, suggesting that the abnormal spermatogenesis observed in the Cnot7(-/-) testes was induced by the impaired testicular microenvironment rather than a germ cell defect. In the present study, we have carried out reciprocal germ cell transplantation in which wild type germ cells were transplanted as donors into the recipient Cnot7(-/-) testes to evaluate the recipient microenvironment for supporting the spermatogenesis of donor cells. We noticed that donor cell colonization was less efficient in Cnot7(-/-) than in Cnot7(-/-) testes, and that the donor derived spermatids in the recipient Cnot7-/- testes showed severe deformities. These results support our previous report that Sertoli cell defects in the Cnot7(+) testes could induce oligo-astheno-teratozoospermia.

DOI： 10.1679/aohc.67.307

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：INT SOC HISTOLOGY & CYTOLOGY  

Spermatogonial stem cells (SSCs), having yet to possess decisive markers, can only be detected retrospectively by transplantation assay. It was reported recently that mouse gonocytes collected from DBA/2 and ICR neonates propagated in vitro. This cultured germ cell, named the germline stem cell (GS cell), produced functional sperm to make progeny when transplanted into recipient mouse testes. Here we show that GS cell lines can be established not only front neonatal testes but also from the testis of adult mice. We also confirmed that GS cells once transplanted into a host testis can be recovered to resume in vitro expansion, indicating that they are convertible mutually with SSCs in adult testes. Confocal laser microscopic examination showed GS cells resemble undifferentiated spermatogonia in the adult testis. This unique cell line could be useful for research in germ cell biology and applicable as a new tool for the genetic engineering of animals.

DOI： 10.1679/aohc.67.297

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Spermatogenesis is a complex process that involves cooperation of germ cells and testicular somatic cells. Various genetic disorders lead to impaired spermatogenesis, defective sperm function and male infertility(1). Here we show that Cnot7(-/-) males are sterile owing to oligo-astheno-teratozoospermia, suggesting that Cnot7, a CCR4-associated transcriptional cofactor(2), is essential for spermatogenesis. Maturation of spermatids is unsynchronized and impaired in seminiferous tubules of Cnot7(-/-) mice. Transplantation of spermatogonial stem cells from male Cnot7(-/-) mice to seminiferous tubules of Kit mutant mice (Kit(W/W-v)) restores spermatogenesis, suggesting that the function of testicular somatic cells is damaged in the Cnot7(-/-) condition. The testicular phenotypes of Cnot7(-/-) mice are similar to those of mice deficient in retinoid X receptor beta (Rxrb)(3). We further show that Cnot7 binds the AF-1 domain of Rxrb and that Rxrb malfunctions in the absence of Cnot7. Therefore, Cnot7 seems to function as a coregulator of Rxrb in testicular somatic cells and is thus involved in spermatogenesis.

DOI： 10.1038/ng1344

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.95.309_4

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We report two cases of XX male with chief complaints of infertility. Physical examination ofboth of both patients aged 42 and 29 demonstrated normal male habitus except for small testes. Semen analyses demonstrated no spermatozoa. Endocrinological examinations showed hypergonadotrophic hypogonadism. Vesiculograms demonstrated normal seminal tracts. Histological examination of their testes did not reveal germ cells
one case lacked seminiferous tubules and there was hyalinization in the seminiferous tubule in another case. Chromosomal analyses of peripheral blood demonstrated 46,XX. The sex-determining region Y gene was positive and DAZ (deleted in azoospermia) gene was negative in both cases.

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A 34-year-old male visited our department with a chief complaint of infertility, hemospermia and decreasing semen volume. Physical examinations were unremarkable and endocrinological tests demonstrated no abnormalities, however, semen analyses showed a seminal volume of 1.0 ml, many red blood cells and a few motile spermatozoa. MR image revealed a midline cystic mass of the prostate and the vesiculogram confirmed the diagnosis as an ejaculatory duct stenosis due to Müllerian duct cyst. We performed transurethral resection of hemi-verumontanum with testicular biopsy and confirmed recanalization of the seminal tract. His testicular histology showed normal spermatogenesis. Since left epididymitis occurred on the 24th day after the surgery, the patient was given 1.0 g/day of Panipenem. Followed by subsequent right epididymitis occurred, his bilateral epididymitis recovered normal consistency after additional dosing of Isepamicin. Postoperative semen analysis showed a seminal volume of 2.5 ml, sperm concentration of 14.0 × 106/ml, sperm motility of 70.0% and no red blood cells. The pregnancy of his spouse was confirmed 7 months after the surgery.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

Recent studies have demonstrated that GnRH-analogues can stimulate regeneration of spermatogenesis of rats when administered after testicular damages. Although the mechanism of this phenomenon has not been elucidated yet, stem cell factor (SCF) produced by Sertoli cells was proposed to mediate the effects of GnRH-analogues on spermatogonial proliferation and/or survival. In the present study, we quantitatively evaluated the proliferation of spermatogonia and addressed whether SCF mediates the effect of GnRH-analogue on spermatogonial proliferation, using a novel approach combining spermatogonial transplantation and laser confocal microscopic observation. In the first experiment, using wild-type mice as recipients for spermatogonial transplantation, the number of donor spermatogonia per 100 Sertoli cells in each spermatogenic colony was significantly higher in the experimental group of mice treated with leuprorelin, a GnRH-agonist, than that of the control group at 4 and 5 wk after transplantation. In the second experiment, Steel/Steel(dickie) (SI/SId) mutant mice, which lack expression of membrane bound form SCIF, were used as recipients. As seen in the first experiment, the number of undifferentiated spermatogonia was significantly higher in leuprorelin-treated than in the control group. Since undifferentiated spermatogonia do not express the receptor of SCF, the present study clearly demonstrates that neither membrane-bound nor secreted forms of SCF are involved in the mechanism of GnRH-analogue's effect on spermatogonial proliferation and/or survival.

DOI： 10.1095/biolreprod.102.013276

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

The stem cell properties of gonocytes and prospermatogonia at prepubertal stages are still largely unknown: it is not clear whether gonocytes and prospermatogonia are a special cell type or similar to adult undifferentiated spermatogonia. To characterize these cells, we have established transgenic mice carrying EGFP (enhanced green fluorescence protein) cDNA under control of an Oct4 18-kb genomic fragment containing the minimal promoter and proximal and distal enhancers; Oct4 is reported to be expressed in undifferentiated spermatogonia at prepubertal stages. Generation of transgenic mice enabled us to purify gonocytes and prospermatogonia from the somatic cells of the testis. Transplantation studies of testicular cells so far have been done with a mixture of germ cells and somatic cells. This is the first report that establishes how to purify germ cells from total testicular cells, enabling evaluation of cell-autonomous repopulating activity of a subpopulation of prospermatogonia. We show that prospermatogonia differ markedly from adult spermatogonia in both the size of the KIT-negative population and cell cycle characteristics. The GFP(+)KIT(-) fraction of prospermatogonia has much higher repopulating activity than does the GFP(+)KIT(+) population in the adult environment. Interestingly, the GFP(+)KIT(+) population still exhibits repopulating activity, unlike adult KIT-positive spermatogonia. We also show that ALCAM, activated leukocyte cell adhesion molecule, is expressed transiently in gonocytes. Sertoli cells and myoid cells also express ALCAM at the same stage, suggesting that ALCAM may contribute to gonocyte-Sertoli cell adhesion and migration of gonoyctes toward the basement membrane. (C) 2003 Elsevier Science (USA). All rights reserved.

DOI： 10.1016/S0012-1606(03)00111-8

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Language：Japanese   Publishing type：Research paper (scientific journal)  

We report a case of bilateral hydronephrosis caused by pseudolymphoma of bilateral renal pelves. A 52-year-old woman with Sjogren's syndrome and bronchial asthma was found to have bilateral hydronephrosis. Abdominal plain computerized tomography showed an irregular thickening of the bilateral renal pelves with moderate hydronephrosis. The gallium scintigraphy revealed intense tracer uptake in bilateral renal pelves. Open biopsy of the right renal pelvis was perfomed under the diagnosis of malignant lymphoma. The pathologic diagnosis was pseudolymphoma of the renal pelvis. Steroid therapy dramatically improved pseudolymphoma and hydronephrosis within a month. There were no signs of recurrence.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

Mammalian male germ cells might be generally thought to have infinite proliferative potential based on their life-long production of huge numbers of sperm. However, there has been little substantial evidence that supports this assumption. In the present study, we performed serial transplantation of spermatogonial stem cells to investigate if they expand by self-renewing division following transplantation. The transgenic mouse carrying the Green fluorescent protein gene was used as the donor cell source that facilitated identification and recollection of colonized donor germ cells in the recipient testes. The established colonies of germ cells in the recipient testes were collected and transplanted to new recipients. This serial transplantation of spermatogonial stem cells repopulated the recipient testes, which were successfully performed sequentially up to four times from one recipient to the next. The incubation periods between two sequential transplantations ranged from 55 to 373 days. During these passages, the spermatogonial stem cells showed constant activity to form spermatogenic colonies in the recipient testis. They continued to increase in number for more than a year following transplantation. Colonization efficiency of spermatogonial stem cells was determined to be 4.25% by using SI/SId mice as recipients that propagated only undifferentiated type A spermatogonia in their testes. Based on the colonization efficiency, one colony-forming activity was assessed to equate to about 20 spermatogonial stem cells. The spermatogonial stem cells were estimated to expand over 50-fold in 100 days in this experiment.

DOI： 10.1095/biolreprod.102.004549

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.94.175_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.94.363_4

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Japanese Urological Association  

(Purpose) we report our experience in 10 years of sperm cryopreservation to reveal the present state of the cryopreservation project. (Materials and Methods) 42 germ cell tumor, 110 non-germ cell tumor and 2 non-malignant disease patients who visited our clinic for semen cryopreservation were retrospectively analyzed. (Results) only 7 (20%) out of 35 unilateral testicular tumor patients who had received no chemotherapy met the WHO criteria for sperm concentration and motility. However, there were no patients with azoospermia. Three testicular tumor patients with previous chemotherapy and 4 retroperitoneal germ cell tumor patients had poor sperm concentrations and motilities. Twenty (52%) out of 38 non-germ cell tumor patients without previous chemotherapy met the WHO criteria. In contrast, only 9 patients (13%) met the WHO criteria among 72 patients with previous chemotherapy. Twenty-nine patients (40%) with chemotherapy were azoospermia. Totally, 74 (96%) of 77 tumor patients' semen without chemotherapy and 34 (45%) of 75 with chemotherapy were cryopreserved. Sperm from a patient with testicular torsion were cryopreserved. (Conclusion) most cancer patients without previous chemotherapy, regardless of underlying disease, had abundant motile sperm. However, half of the patients who had received chemotherapy did not have suitable sperm for freezing. It is important to inform young cancer patients of the cryopreservation project immediately after the diagnosis is made.

DOI： 10.5980/jpnjurol1989.94.513

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.94.173_1

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DOI： 10.5980/jpnjurol.94.269_2

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：LIPPINCOTT WILLIAMS & WILKINS  

Purpose: Cryptorchidism is an adverse condition of spermatogenesis in many mammals. Surgical cryptorchidism in rats lasting more than a few weeks is so detrimental that spermatogenesis cannot be completely recovered even after orchiopexy. We evaluated the efficacy of the high dose gonadotropin-releasing hormone (Gn-RH) agonist leuprorelin acetate on damaged spermatogenesis in rat cryptorchid testes.
Materials and Methods: Male Fisher rats were divided into 2 groups of 6 each and bilateral cryptorchidism was artificially produced. Five weeks later all rats underwent bilateral orchiopexy. One group served as the control, while the other received Gn-RH agonist injections at orchiopexy and 4 weeks later. The animals were sacrificed 15 weeks after orchiopexy. The weight of the body, testis and epididymis was measured and the histology of spermatogenesis was examined. For statistical analysis the Student t test was applied.
Results: Testes in the Gn-RH group rats showed significant recovery of spermatogenesis up to complete spermatozoa formation, while those in control rats remained almost degenerated. The mean incidence of seminiferous tubules with recovered spermatogenesis plus or minus standard deviation was significantly higher in the Gn-RH than in the control group (87.8% +/- 6.0% versus 12.5% +/- 7.7%, p &lt; 0.001).
Conclusions: Administering the high dose Gn-RH agonist leuprorelin acetate after orchiopexy greatly enhanced the recovery of spermatogenesis in rats. This finding is in accordance with other recent reports that treatment with Gn-RH analogues promotes the regeneration of once damaged spermatogenesis. On the other hand, these findings may cause one to question supplementation therapy now used in regular practice to boys with cryptorchidism.

DOI： 10.1097/01.ju.0000024626.25898.0f

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Japanese Urological Association  

(Purpose) To assess the effect of high concentration of sodium and potassium ion on the motilities of human sperm following the preservation in electrolyte-free solution. (Patient) Semen samples were obtained from patients attending our infertility clinic. The motilities of their sperm were more than 40%. (Method) Ejaculated sperm were washed by using the electrolyte-free Percoll gradient and then preserved in electrolyte-free solution at 4 degrees C for a week, The preserved sperm was incubated and analyzed after the addition of electrolyte solutions under acid or alkaline condition, The sperm were continuously preserved for a day at room temperature and analyzed, (Result) The sperm was by the addition of 35∼135mM Na+ or 5∼135mM K+ in acid solutions. Under the alkaline conditions, sperm were reactivated without Na+ or K+. However, the addition of 135mM Na+ or K+ significantly decreased in sperm motilities. The some sperm in the solution containing low concentration of Na+ or K+ ions maintained their motilities for a day at room temperature. However, all sperm lost their motilities in the 135mM Na+ or K- solutions, (Conclusion) The high concentration of Na+ or K+ ions had detrimental effects on motilities of sperm following the preservation in electrolyte-free solution.

DOI： 10.5980/jpnjurol1989.93.440

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.93.204_1

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DOI： 10.5980/jpnjurol.93.206_2

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：BLACKWELL PUBLISHING ASIA  

Background: Future fertility is a major concern for cancer patients who undergo intensive chemotherapy. There has been controversy about whether hormonal treatments may have protective effects against the severe spermatogenic damage caused by chemotherapy or irradiation. Recently, it has been proposed that gonadotrophin-releasing hormone (GnRH) analogs administered after testicular damage stimulate the recovery of spermatogenesis. In this study, we have investigated the effects of GnRH agonist, leuprorelin, on the damage to spermatogenesis induced by busulfan.
Methods: Fisher rats were treated with busulfan, 25 mg/kg, intraperitoneally. The effects of subcutaneous injections of leuprorelin before or after treatment were evaluated histologically 18 weeks later.
Results: The percentage of 'recovered' seminiferous tubules was 27.7 +/- 12.6% in control rats without leuprorelin and 26.9 +/- 10.2% in rats with leuprorelin injected 4 weeks before busulfan. Rats in both groups showed poor recovery of spermatogenesis with an increase of intratesticular fluid. However. rats treated with leuprorelin three times (4 weeks apart) after busulfan showed an improvement of up to 56.5 +/- 12.0% (P &lt; 0.05). A focal but massive necrotic lesion in the testis was observed only in this group of rats.
Conclusions: The results demonstrated that leuprorelin administered after chemical testicular damage enhanced the recovery of spermatogenesis. At the same time, a possible significant side-effect of leuprorelin was noted.

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Language：English   Publisher：SPRINGER-VERLAG  

DOI： 10.1007/s001090100228

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：CHURCHILL LIVINGSTONE  

Gonadotropin-releasing hormone (GnRH)-agonist or antagonist treatment supports recovery of spermatogenesis after irradiation damage in the rat and appears to be beneficial to colonization of recipient testes after spermatogonial transplantation from fertile donors to the testes of infertile recipients in rats and mice. in the present study, we quantified the effect of treatment of recipient mice with the GnRH-agonist leuprolide acetate on the extent of colonization by donor spermatogonial stern cells in the recipient testis. Testis cells from mice carrying transgenes, which produce beta -galactosidase in spermatogenic cells, were used as donor cells for transplantation to allow for quantification of donor spermatogenesis in the recipient testis by staining for enzyme activity. Donor cell colonization 3 months after transplantation was compared between recipients receiving leuprolide in different treatment protocols and untreated control mise. Two injections of leuprotide 4 weeks apart prior to transplantation with as little as 3.8 mg/kg resulted in a pronounced improvement in the number of donor-derived spermatogenic colonies as well as in the in the area of recipient seminiferous tubules occupied by donor cell spermatogenesis, improved colonization efficiency by treatment with GnRH-agonist can make the technique of spermatogonial transplantation applicable to situations when only low numbers of donor cells are available. (C) 2001 Harcourt Publishers Ltd.

DOI： 10.1054/tice.2001.0177

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.92.292_2

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.92.410_1

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.92.216_2

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Language：Japanese   Publisher：(一社)日本移植学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE AMERICA INC  

Azoospermia or oligozoospermia due to disruption of spermatogenesis are common causes of human male infertility. We used the technique of spermatogonial transplantation in two infertile mouse strains, Steel (SI) and dominant white spotting (W), to determine if stem cells from an infertile male were capable of generating spermatogenesis. Transplantation of germ cells from infertile SI/SId mutant male mice to infertile W/W-v or W-v/W-54 mutant male mice restored fertility to the recipient mice. Thus, transplantation of spermatogonial stem cells from an infertile donor to a permissive testicular environment can restore fertility and result in progeny with the genetic makeup of the infertile donor male.

DOI： 10.1038/71496

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Language：Japanese   Publisher：一般社団法人 日本泌尿器科学会  

DOI： 10.5980/jpnjurol.91.366_3

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DOI： 10.5980/jpnjurol.91.364_4

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Transplantation of spermatogonial stem cells from fertile, transgenic donor mice to the testes of infertile recipients provides a unique system to study the biology of spermatogonial stem cells. To facilitate the investigation of treatment effects on colonization efficiency an analysis system was needed to quantify colonization of recipient mouse seminiferous tubules by donor stem cell-derived spermatogenesis. In this study, a computer-assisted morphometry system was developed and validated to analyze large numbers of samples. Donor spermatogenesis in recipient testes is identified by blue staining of donor-derived spermatogenic cells expressing the E. coli lacZ structural gene. Images of seminiferous tubules from recipient testes collected three months after spermatogonial transplantation are captured, and stained seminiferous tubules containing donor-derived spermatogenesis are selected for measurement based on their color by color thresholding. Colonization is measured as number, area, and length of stained tubules. Interactive, operator-controlled color selection and sample preparation accounted for less than 10% variability for all collected parameters. Using this system, the relationship between number of transplanted cells and colonization efficiency was investigated. Transplantation of 10(4) cells per testis only rarely resulted in colonization, whereas after transplantation of 10(5) and 10(6) cells per testis the extent of donor-derived spermatogenesis was directly related to the number of transplanted donor cells. It appears that about 10% of transplanted spermatogonial stem cells result in colony formation in the recipient testis. The present study establishes a rapid, repeatable, semi-interactive morphometry system to investigate treatment effects on colonization efficiency after spermatogonial transplantation in the mouse. Mel. Reprod. Dev. 53:142-148, 1999. (C) 1999 Wiley-Liss, Inc.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC STUDY REPRODUCTION  

It was recently demonstrated that rat spermatogenesis can occur in the seminiferous tubules of an immunodeficient recipient mouse after transplantation of testis cells from a donor rat, In the present study, hamster donor testis cells were transplanted to mice to determine whether xenogeneic spermatogenesis would result, The hamster diverged at least 16 million years ago from the mouse and produces spermatozoa that are larger than, and have a shape distinctly different from, those of the mouse, In four separate experiments with a total of 13 recipient mice, hamster spermatogenesis was identified in the testes of each mouse. Approximately 6% of the tubules examined demonstrated xenogeneic spermatogenesis, In addition, cryopreserved hamster testis cells generated spermatogenesis in recipients. However, abnormalities were noted in hamster spermatids and acrosomes in seminiferous tubules of recipient mice. Hamster spermatozoa were also found in the epididymis of recipient animals, but these spermatozoa generally lacked acrosomes, and heads and tails were separated. Thus, defects in spermiogenesis occur in hamster spermatogenesis in the mouse, which may reflect a limited ability of endogenous mouse Sertoli cells to support fully the larger and evolutionarily distant hamster germ cell. The generation of spermatogenesis from frozen hamster cells now adds this species to the mouse and rat, in which spermatogonial stem cells also can be cryopreserved. This finding has immediate application to valuable animals of many species, because the cells could be stored until suitable recipients are identified or culture techniques devised to expand the stem cell population.

DOI： 10.1095/biolreprod60.2.515

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Language：English   Publishing type：Research paper (scientific journal)  

Objective: To assess the viability and function of human sperm in electrolyte-free cold preservation. Design: Prospective comparative study. Setting: Andrology laboratory of our hospital. Patient(s): Ten semen samples obtained from patients attending our infertility clinic. Intervention(s): Ejaculated sperm were washed using the electrolyte-free Percoll gradient and were then preserved in 0.33 M glucose solution, 0.16 M NaCl solution, 0.16 M KCl solution at 4°C for 4 weeks. As a control, TEST (TES and Tris) yolk buffer (TYB) was added to the ejaculated semen and preserved at 4°C. Main Outcome Measure(s): Sperm tail morphology, motility, viability (eosin-Y stain), and the concentration of adenosine triphosphate (ATP) were analyzed. Result(s): The number of sperm with normal tail form and the motility of sperm preserved in glucose solution (electrolyte-free cold preservation) were significantly (P &lt
0.01) higher for 4 weeks than those of sperm preserved in the other three media. The sperm viability in glucose solution was 75.5%, 65.4%, and 51.3%, after 1, 2, and 4 weeks, respectively. The ATP concentration after 1, 2, and 4 weeks remained 64.2%, 53.0%, and 4.3% of the prestorage value, respectively, in the sperm stored in glucose solution. Conclusion(s): The morphology, motility, viability, and ATP concentration of sperm in electrolyte-free cold preservation were substantially better than those in NaCl solution, KCl solution, or TYB for 2 weeks.

DOI： 10.1016/S0015-0282(97)00439-1

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DOI： 10.11477/mf.1409208497

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Language：English   Publishing type：Rapid communication, short report, research note, etc. (scientific journal)  

DOI： 10.1016/j.urology.2012.08.017

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Language：English   Publishing type：Rapid communication, short report, research note, etc. (scientific journal)   Publisher：ELSEVIER SCIENCE INC  

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DOI： 10.1089/end.2012.1528

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Language：English   Publishing type：Rapid communication, short report, research note, etc. (scientific journal)   Publisher：WILEY-BLACKWELL  

DOI： 10.1111/j.1464-410X.2012.11000_2.x

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Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：Japanese   Publisher：学研メディカル秀潤社  

CiNii Books

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Other Link： http://search.jamas.or.jp/link/ui/2011302606

Language：English   Publishing type：Book review, literature introduction, etc.   Publisher：John Wiley and Sons Ltd  

Spermatogonial stem cells (SSCs) provide the basis for the life-long production of enormous numbers of sperm. The nature of these mysterious cells is being clarified. Although they were regarded to be mostly dormant, dividing rarely and remaining static in a niche, their rather dynamic behavior in the seminiferous tubules has been disclosed. The territories of each colony of SSCs can also quickly change in size. The development of a culture method for SSCs also shed light on their stable, but at the same time, fragile characteristics. In addition, an in vitro system for spermatogenesis was developed which can produce functional sperm from SSCs. These new developments will contribute to reproductive medicine. © 2011 Japan Society for Reproductive Medicine.

DOI： 10.1007/s12522-011-0084-7

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Language：Japanese   Publisher：日本生殖内分泌学会  

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Other Link： http://search.jamas.or.jp/link/ui/2011081402

Language：Japanese   Publisher：(一社)日本生殖医学会  

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：SOC STUDY REPRODUCTION  

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Language：Japanese   Publisher：泌尿器科紀要刊行会  

78歳,女。肉眼的血尿で受診し、膀胱後壁に4cm大の有茎性乳頭状腫瘍を認め1ヵ月後TUR-Btを施行し、病理所見で移行性上皮癌、G3、pT1であった。肉眼的には完全に切除できたと考えたが腫瘍径が大きいことや病理所見を考え再発予防目的でBCG膀胱内注入療法を8回施行した。術後1年10ヵ月に不正性器出血が出現し、子宮体部にT2、high intensityな腫瘤を認め、経腟的子宮内腫瘍生検により移行上皮癌G2で膀胱子宮転移と診断した。年齢、本人の希望により放射線療法単独(腟内照射+全骨盤外照射60Gy)による治療を行い照射後4ヵ月経過して、膀胱内再発はなく子宮内腫瘤も増大を認めていない。

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Language：Japanese   Publisher：(一社)日本癌治療学会  

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：MARY ANN LIEBERT INC  

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Language：Japanese   Publisher：神奈川県医師会  

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Language：Japanese   Publisher：医学図書出版(株)  

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Language：English   Publishing type：Book review, literature introduction, etc.   Publisher：John Wiley and Sons Ltd  

Detection of spermatogonial stem cells (SSC) became possible 10 years ago, with the transplantation of germ cells into the seminiferous tubules of mice. Using this assay system, attempts to maintain and expand SSC in vitro finally bore fruit. Gonocytes from neonatal mice and spermatogonial stem cells from adult mice were plated on feeder cells in a medium supplemented with Glial cell line-derived neurotrophic factor (GDNF) along with certain other factors. The germ cells that emerged under such conditions, named germline stem (GS) cells, proliferated exponentially through self-renewing division. GS cells in vitro show features of differentiation as well. Some expressed c-kit, which is a cell surface marker of differentiating spermatogonia. Therefore, it seems that GS cells undergo both self-renewing and differentiating cell divisions in vitro. There is a century of history behind attempts to reproduce spermatogenesis in vitro and significant progress has been made. Nonetheless, there are few established culture-based protocols for recreating spermatogenesis in vitro. GS cells would be an ideal starting material in this regard. (Reprod Med Biol 2006
5: 169-174): © 2006 The Authors Journal compilation 2006 Japan Society for Reproductive Medicine.

DOI： 10.1111/j.1447-0578.2006.00138.x

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：SOC STUDY REPRODUCTION  

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Other Link： http://search.jamas.or.jp/link/ui/2004041595

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Other Link： http://search.jamas.or.jp/link/ui/2004041589

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Other Link： http://search.jamas.or.jp/link/ui/2004041596

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Other Link： http://search.jamas.or.jp/link/ui/2001061907

Language：English   Publisher：CHURCHILL LIVINGSTONE  

Testis cell transplantation from mice or rats into recipient mouse seminiferous tubules results in donor cell-derived spermatogenesis in nearly all host testes. Normal spermatozoa are produced and, in the most successful mouse transplantations, the donor haplotype is transmitted to progeny of the recipient. However, few studies have been performed in other species. In this report, we demonstrate that rat and mouse testis cells will generate donor cell-derived spermatogenesis in recipient rat seminiferous tubules. Depletion of endogenous spermatogenesis before donor cell transplantation was more difficult in rat than reported for mouse recipients. A protocol employing treatment of neonatal rats with busulfan was most effective in preparing recipients and allowed more than 90% of testes to be colonized by donor cells. Transplantation of mouse testis cells into rat seminiferous tubules was most successful in recipients made cryptorchid and treated with busulfan. In the best experiments, about 55% of rat testes were colonized by mouse cells. Both rat and mouse donor cell-derived spermatogenesis were improved by treatment of rat recipients with leuprolide, a gonadotropin-releasing hormone agonist. The studies indicated that recipient preparation for spermatogonial stem cell transplantation was critical in the rat and differs from the mouse. However, modification of currently used techniques should allow male germ line stem cell transplantation in many species. (C) 1999 Harcourt Publishers Ltd.

DOI： 10.1054/tice.1999.0060

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Language：English   Publisher：WILEY-BLACKWELL  

Transplantation of spermatogonial stem cells from fertile, transgenic donor mice to the testes of infertile recipients provides a unique system to study the biology of spermatogonial stem cells. To facilitate the investigation of treatment effects on colonization efficiency an analysis system was needed to quantify colonization of recipient mouse seminiferous tubules by donor stem cell-derived spermatogenesis. In this study, a computer-assisted morphometry system was developed and validated to analyze large numbers of samples. Donor spermatogenesis in recipient testes is identified by blue staining of donor-derived spermatogenic cells expressing the E. coli lacZ structural gene. Images of seminiferous tubules from recipient testes collected three months after spermatogonial transplantation are captured, and stained seminiferous tubules containing donor-derived spermatogenesis are selected for measurement based on their color by color thresholding. Colonization is measured as number, area, and length of stained tubules. Interactive, operator-controlled color selection and sample preparation accounted for less than 10% variability for all collected parameters. Using this system, the relationship between number of transplanted cells and colonization efficiency was investigated. Transplantation of 10(4) cells per testis only rarely resulted in colonization, whereas after transplantation of 10(5) and 10(6) cells per testis the extent of donor-derived spermatogenesis was directly related to the number of transplanted donor cells. It appears that about 10% of transplanted spermatogonial stem cells result in colony formation in the recipient testis. The present study establishes a rapid, repeatable, semi-interactive morphometry system to investigate treatment effects on colonization efficiency after spermatogonial transplantation in the mouse. Mel. Reprod. Dev. 53:142-148, 1999. (C) 1999 Wiley-Liss, Inc.

DOI： 10.1002/(SICI)1098-2795(199906)53:2<142::AID-MRD3>3.0.CO;2-O

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Language：English   Publisher：CHURCHILL LIVINGSTONE  

Mouse-to-mouse transplants were studied at 10 min, 9 h, 24 h, 1 week, 1 month, 2 months, and 3 months post-transplantation. Data from a previous light microscope study were confirmed and extended using morphometric and ultrastructural techniques. As soon as 10 min after introduction of the germ cells from one mouse into the tubule lumen of a recipient mouse they developed relationships with small Sertoli cell processes. The extent of this surface-to-surface relationship increased in animals sacrificed up to 1 week post-transplantation, Most transplanted germ cells retained the characteristics of the donor germ cells after they had been isolated and pelleted, Nearly all transplanted cells eventually underwent phagocytosis by the recipient Sertoli cells. The presence of small apparent clones of germ cells after 1 week of transplantation indicated that some germ cells may divide and survive for short periods within the epithelium. No discernible qualitative subcellular changes in the host Sertoli cell accompanying the development of transplant spermatogenesis were noted. Macrophages were present in the region of the boundary tissue between myoid cells and appeared to increase in number in the peritubular tissue of transplanted testes. Images suggest that they migrated into the tubule to gain entrance to the lumen and there take on the form of activated macrophages. Some macrophages phagocytose sperm at 2 months and 3 months post-transplantation. A testis weight increase previously demonstrate to occur at 24 h post-introduction of germ cells was found to be due to an increase in the volume of the tubular lumen. The increase of lumen size at 24 h was not related to the volume of the injected material. It is suggested that the presence of injected cells, likely germ cells, in the tubule lumen stimulated increased secretion by the Sertoli cell.

DOI： 10.1054/tice.1999.0006

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Language：English   Publisher：SOC STUDY REPRODUCTION  

Spermatogenesis is one of the most productive self-renewing systems in the body: on the order of 10(7) spermatozoa are produced daily per gram of testis tissue. In each mammalian species, the time required for completion of the process is unique and unalterable. Because the process is supported by somatic Sertoli cells, it has generally been thought that cell-cell interaction between germ and Sertoli cells controls the duration of cell cycles and cellular organization. We have used the newly developed technique of spermatogonial transplantation to examine which cell type(s) determines the rate at which germ cells proceed through spermatogenesis. Rat germ cells were trans-planted into a mouse testis, and the mouse was killed 12.9-13 days after administration of a single dose of [H-3]thymidine. The most advanced rat cell type labeled was the pachytene spermatocyte at stages VI-VIII of the spermatogenic cycle. In animals given only rat cells, some endogenous spermatogenesis of the mouse recovered. The most advanced labeled mouse cell types in recipients killed 12.9-13 days after administration of a single dose of [H-3]thymidine were meiotic cells or young spermatids, which is consistent with a spermatogenic cycle length comparable to the 8.6 days reported for the mouse. The same results were obtained if a mixture of rat and mouse cells were transplanted. There existed two separate timing regimens for germ cell development in the recipient mouse testis; one of rat and one of mouse duration. Rat germ cells that were supported by mouse Sertoli cells always differentiated with cell cycle timing characteristic of the rat and generated the spermatogenic structural pattern of the rat, demonstrating that the cell differentiation process of spermatogenesis is regulated by germ cells alone.

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Language：English   Publisher：SOC STUDY REPRODUCTION  

Development of spermatogonial transplants was studied by using 5- to 6-wk-old histocompatible mice as cell donors and sterile (W-locus) mice as recipients. Groups of animals transplanted with germ cell suspensions were killed at 10 min, 9 h, 24 hi 1 wk, 1 mo, 2 mo, and 3 mo along with age-matched "start" and "end" W-locus controls. Weight of testes increased significantly at 24 h through 3 mo after germ cell transplantation, suggesting that the infused cells quickly stimulated organ function, Small clones of young spermatocytes were evident at 1 mo and sperm at 2 mo. The percentage of tubular profiles containing active spermatogenesis originating from spermatogonia increased with time (0.8% at 1 mo, 8.9% at 2 mo, and 28.2% at 3 mo). Most transplanted germ cells were eliminated from the seminiferous epithelium through phagocytosis by Sertoli cells that occurred primarily before `1 wk, although some pachytene cells were able to proceed through meiosis by 1 wk, A variety of abnormal features are described that characterize developing spermatogenesis in the transplanted testis, Spermatogenesis improved quantitatively and qualitatively with time although released sperm were frequently engulfed by intratubular macrophages and Sertoli cells. A quantitative analysis of spermatogenesis from transplanted germ cells will serve as a basis for improving spermatogonial transplant efficiency.

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Language：English   Publisher：MARY ANN LIEBERT INC PUBL  

We previously identified the ZNF85 (HPF4) KRAB zinc finger gene, a member of the human ZNF91 family. Here, we show that the ZNF85 gene is highly expressed in normal adult testis, in seminomas, and in the NT2/D1 teratocarcinoma cell line. Immunocytochemical localization of a panel of beta-Gal/ZNF85 fusion proteins revealed that ZNF85 contains at least one nuclear localization signal located in the spacer region connecting the KRAB domain with the zinc finger repeats. Bacterially expressed ZNF85 zinc finger domain bound strongly and exclusively to DNA in vitro in a zinc-dependent manner. The KRAB(A) domain of the ZNF85 protein and of several other members of the ZNF91 family exhibited repressing activity when tested in Gal4 fusion protein assays. The repression was significantly enhanced by the addition of the KRAB (B) domain, whereas further addition of other conserved regions had no effect. The ZNF85 KRAB(A) and (B) domains in vitro bound several nuclear proteins that might constitute critical cofactors for repression.

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Language：English   Publisher：CHURCHILL LIVINGSTONE  

Spermatogonial stem cells can be transplanted from a fertile donor mouse to the testis of an infertile recipient where they establish spermatogenesis and produce spermatozoa, In the present study we investigated whether treatment of recipient mice with the gonadotropin-releasing hormone (GnRH) agonist leuprolide acetate could alter the efficiency of colonization by donor spermatogonial stem cells in the recipient testis. Six recipient mice were treated with busulfan to destroy endogenous spermatogenesis followed by injection of leuprolide acetate to three of the mice. Testis cells from mice carrying the ZFlacZ transgene, which produces beta-galactosidase in spermatids, were used as donor cells for transplantation to allow for identification of donor spermatogenesis in the recipient testis by staining for enzyme activity. The extent of donor cell colonization was compared between leuprolide treated recipients and untreated control mice 3 months after transplantation. Efficiency of colonization by donor cells was markedly enhanced in recipient mice treated with the GnRH agonist leuprolide acetate, which makes the technique of spermatogonial transplantation applicable to a wide range of experimental situations. The present study also indicates that this technique can be used as a biological assay system to investigate factors controlling the establishment and progression of spermatogenesis.

DOI： 10.1016/S0040-8166(98)80039-6

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Language：English   Publisher：ELSEVIER SCIENCE INC  

Six Kruppel-type zinc finger (ZF) genes were cloned from a seminoma cDNA library. One, ZFS-1, showed high sequence homology to the ZNF91 KRAB [Kruppel-associated box] ZF gene family and also the same chromosomal assignment. Interestingly, Northern blot analyses using ZFS-1 and ZNF91 revealed that multiple ZF genes on chromosome 19 were predominantly expressed in seminomas. In addition, the testis and the seminoma showed specific expression of 2.3 kb transcript. Our results suggest that ZF genes on chromosome 29 may be implicated in the development and/of growth of seminomas. (C) Elsevier Science Inc., 1998.

DOI： 10.1016/S0165-4608(97)00004-6

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：SOC STUDY REPRODUCTION  

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Language：English   Publisher：UNIV BASQUE COUNTRY PRESS  

In the adult male, germ cell differentiation takes place in the seminiferous tubules of the testis by a complex, highly organized and very efficient process. A population of diploid stem-cell spermatogonia that lie on the basement membrane of the tubule continuously undergoes self-renewal and produces progeny cells, which initiate the process of cellular differentiation to generate mature spermatozoa. Each testis contains many seminiferous tubules, which are connected at both ends to a collecting system called the rete testis. The mature spermatozoa pass from the tubules into the rete and are then carried through efferent ducts to the epididymis for final maturation before they are ready to fertilize an egg. In previous studies, we have demonstrated that donor testis cells collected from a fertile mouse are able to generate spermatogenesis when transplanted to the seminiferous tubules of an infertile male. The spermatozoa produced by the recipient from the donor-derived spermatogonial stem cells are able to fertilize eggs and produce progeny carrying the donor male haplotype, Furthermore, donor testis stem cells from a rat will generate normal rat spermatozoa following transplantation to a mouse testis. The spermatogonial transplantation technique is clearly valuable and applicable to many species, but it is difficult, Therefore, several procedures to introduce donor cells into the seminiferous tubules of a recipient have been developed using the mouse as a model, and they are described here in detail. The results indicate that microinjection of cell suspensions into the seminiferous tubules, efferent ducts or rete testis are equally effective in generating donor cell-derived spermatogenesis in recipients, Each approach is likely to be useful for different experimental purposes in a variety of species.

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Language：English   Publisher：SOC TOXICOLOGIC PATHOLOGISTS  

This study discusses mice bronchiolar epithelium following bromobenzene (BrBZ) exposure. Our study shows 3 cell types following BrBZ exposure: Clara cells, ciliated cells, and newly discovered BrBZ-resistant nonciliated cells. Through electron microscopic autoradiography, we were able to show the proliferative ability of BrBZ-resistant nonciliated cells. This finding suggests that BrBZ-resistant nonciliated cells may function as progenitor cells. In addition to discovering this new cell type, our results also demonstrate and confirm the proliferative ability of modified Clara cells.

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