In vitro production of haploid cells after coculture of CD49f+ with Sertoli cells from testicular sperm extraction in nonobstructive azoospermic patients

Riboldi, M.; Rubio, Carmen; Pellicer, António; Gil-Salom, Manuel; Simón, Carlos

doi:10.1016/j.fertnstert.2012.05.039

Cited by 36 publications

(36 citation statements)

References 46 publications

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“…After a 5‐day incubation, the immunostaining of SCP3 and CREST showed the existence of the meiotic germ cells. FISH analysis confirmed the appearance of the haploid cells . In this study again, 5 days are too short a period to complete such a complicated process of differentiation.…”

Section: Cell Culture Method Two Dimensional (2d)supporting

confidence: 67%

In vitro spermatogenesis: A century‐long research journey, still half way around

Komeya

Sato

Ogawa

2018

Reprod Medicine & Biology

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BackgroundSpermatogenesis is one of the most complicated cellular differentiation processes in a body. Researchers struggled to find and develop a micro‐environmental condition that can support the process in vitro. Such endeavors can be traced back to a century ago and are yet continuing.MethodsReports on in vitro spermatogenesis and related works were selected and classified into four categories based on the method used; organ culture, tubule culture, cell culture, and 3‐dimensional cell culture methods. Each report was critically reviewed from the present point of view by authors who have been working on in vitro spermatogenesis with organ culture method over a decade.ResultsThe organ culture method has the longest history and is the most successful method, which produced fertile mouse sperm from spermatogonial stem cells. Formulation of the medium was a key factor, most importantly serum‐derived substances. However, factors in the serum that induce and support spermatogenesis in the cultured tissue remain to be identified. In addition, the success of mouse spermatogenesis is yet to be applied to other animals. On looking into the history of cell culture method, it became clear that Sertoli cells as feeder cells play an important role. Even with Sertoli cells, however, spermatogenic development has been limited to small parts of spermatogenesis, a segmented period of meiotic prophase for instance. Recent developments of organoid or 3‐dimensional culture techniques are promising but they still need further refinements.ConclusionThe study of in vitro spermatogenesis progressed significantly over the last century. We need more work, however, to establish a culture system that can induce and maintain complete spermatogenesis of many if not all mammalian species.

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Section: Cell Culture Method Two Dimensional (2d)supporting

confidence: 67%

In vitro spermatogenesis: A century‐long research journey, still half way around

Komeya

Sato

Ogawa

2018

Reprod Medicine & Biology

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“…Our dairy goat mGSCs maintained typical GSC and ES‐like colonies when cultured under optimum culture conditions for 9 days ; cells expressed PLZF, VASA , OCT4 (also known as POU5F1), NANOS2 , NANOG , LIN28 and CD49F as analysed by immunofluorescence (Fig. ).…”

Section: Resultsmentioning

confidence: 86%

Promyelocytic leukaemia zinc finger maintains self‐renewal of male germline stem cells (mGSCs) and its expression pattern in dairy goat testis

Song

Zhu

et al. 2013

Cell Proliferation

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“…In addition, RA is known to support the differentiation of pluripotent cells into male germ cell lineage in vitro (Miryounesi et al ., 2014; Zhang et al ., 2015), and also the differentiation of male germ cells into meiotic cells in vitro (Riboldi et al ., 2012; Travers et al ., 2013). The concentration of retinoic acid used here in this study was 10 −6 M, which was found in the previous studies to support germ cell differentiation in vitro (Travers et al ., 2013; Zhang et al ., 2015).…”

Section: Discussionmentioning

confidence: 99%

In vitrodifferentiation of rat spermatogonia into round spermatids in tissue culture

et al. 2016

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STUDY QUESTIONDo the organ culture conditions, previously defined for in vitro murine male germ cell differentiation, also result in differentiation of rat spermatogonia into post-meiotic germ cells exhibiting specific markers for haploid germ cells?SUMMARY ANSWERWe demonstrated the differentiation of rat spermatogonia into post-meiotic cells in vitro, with emphasis on exhibiting, protein markers described for round spermatids.WHAT IS KNOWN ALREADYFull spermatogenesis in vitro from immature germ cells using an organ culture technique in mice was first reported 5 years ago. However, no studies reporting the differentiation of rat spermatogonia into post-meiotic germ cells exhibiting the characteristic protein expression profile or into functional sperm have been reported.STUDY DESIGN, SAMPLES/MATERIALS, METHODSOrgan culture of testicular fragments of 5 days postpartum (dpp) neonatal rats was performed for up to 52 days. Evaluation of microscopic morphology, testosterone levels, mRNA and protein expression as measured by RT-qPCR and immunostaining were conducted to monitor germ cell differentiation in vitro. Potential effects of melatonin, Glutamax® medium, retinoic acid and the presence of epidydimal fat tissue on the spermatogenic process were evaluated. A minimum of three biological replicates were performed for all experiments presented in this study. One-way ANOVA, ANOVA on ranks and student's t-test were applied to perform the statistical analysis.MAIN RESULTS AND THE ROLE OF CHANCEMale germ cells, present in testicular tissue pieces grown from 5 dpp rats, exhibited positive protein expression for Acrosin and Crem (cAMP (cyclic adenosine mono phosphate) response element modulator) after 52 days of culture in vitro. Intra-testicular testosterone production could be observed after 3 days of culture, while when epididymal fat tissue was added, spontaneous contractility of cultured seminiferous tubules could be observed after 21 days. However, no supportive effect of the supplementation with any factor or the co-culturing with epididymal fat tissue on germ cell differentiation in vitro or testosterone production was observed.LIMITATIONS, REASONS FOR CAUTIONThe human testis is very different in physiology from the rat testis, further investigations are still needed to optimize the organ culture system for future use in humans.WIDER IMPLICATIONS OF THE FINDINGSThe successful differentiation of undifferentiated spermatogonia using the testis explant culture system might be employed in future to produce sperm from human spermatogonia as a clinical tool for fertility preservation in boys and men suffering infertility.LARGE SCALE DATANone.STUDY FUNDING AND COMPETING INTEREST(S)This work was supported financially by the Frimurare Barnhuset in Stockholm, the Paediatric Research Foundation, Jeanssons Foundation, Sällskåpet Barnåvard in Stockholm, Swedish Research Council/Academy of Finland, Emil and Wera Cornells Foundation, Samariten Foundation, the Swedish Childhood Cancer Foundation as well as through the regional agre...

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In vitro production of haploid cells after coculture of CD49f+ with Sertoli cells from testicular sperm extraction in nonobstructive azoospermic patients

Cited by 36 publications

References 46 publications

In vitro spermatogenesis: A century‐long research journey, still half way around

In vitro spermatogenesis: A century‐long research journey, still half way around

Promyelocytic leukaemia zinc finger maintains self‐renewal of male germline stem cells (mGSCs) and its expression pattern in dairy goat testis

In vitrodifferentiation of rat spermatogonia into round spermatids in tissue culture

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