Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Differentiation of human male germ cells from Wharton's jelly-derived mesenchymal stem cells

  • Dissanayake, DMAB (Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Kelaniya) ;
  • Patel, H (StemCure Pvt. Ltd.) ;
  • Wijesinghe, PS (Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Kelaniya)
  • Received : 2017.10.01
  • Accepted : 2018.05.03
  • Published : 2018.06.30
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Abstract

Objective: Recapitulation of the spermatogenesis process in vitro is a tool for studying the biology of germ cells, and may lead to promising therapeutic strategies in the future. In this study, we attempted to transdifferentiate Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) into male germ cells using all-trans retinoic acid and Sertoli cell-conditioned medium. Methods: Human WJ-MSCs were propagated by the explant culture method, and cells at the second passage were induced with differentiation medium containing all-trans retinoic acid for 2 weeks. Putative germ cells were cultured with Sertoli cell-conditioned medium at $36^{\circ}C$ for 3 more weeks. Results: The gene expression profile was consistent with the stage-specific development of germ cells. The expression of Oct4 and Plzf (early germ cell markers) was diminished, while Stra8 (a premeiotic marker), Scp3 (a meiotic marker), and Acr and Prm1 (postmeiotic markers) were upregulated during the induction period. In morphological studies, approximately 5% of the cells were secondary spermatocytes that had completed two stages of acrosome formation (the Golgi phase and the cap phase). A few spermatid-like cells that had undergone the initial stage of tail formation were also noted. Conclusion: Human WJ-MSCs can be transdifferentiated into more advanced stages of germ cells by a simple two-step induction protocol using retinoic acid and Sertoli cell-conditioned medium.

Keywords

References

  1. Bellve AR, Cavicchia JC, Millette CF, O'Brien DA, Bhatnagar YM, Dym M. Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization. J Cell Biol 1977;74:68-85. https://doi.org/10.1083/jcb.74.1.68
  2. Clark AT, Bodnar MS, Fox M, Rodriquez RT, Abeyta MJ, Firpo MT, et al. Spontaneous differentiation of germ cells from human embryonic stem cells in vitro. Hum Mol Genet 2004;13:727-39. https://doi.org/10.1093/hmg/ddh088
  3. Petit FG, Kervarrec C, Jamin SP, Smagulova F, Hao C, Becker E, et al. Combining RNA and protein profiling data with network interactions identifies genes associated with spermatogenesis in mouse and human. Biol Reprod 2015;92:71.
  4. Lee DR, Kaproth MT, Parks JE. In vitro production of haploid germ cells from fresh or frozen-thawed testicular cells of neonatal bulls. Biol Reprod 2001;65:873-8. https://doi.org/10.1095/biolreprod65.3.873
  5. Nayernia K, Nolte J, Michelmann HW, Lee JH, Rathsack K, Drusenheimer N, et al. In vitro-differentiated embryonic stem cells give rise to male gametes that can generate offspring mice. Dev Cell 2006;11:125-32. https://doi.org/10.1016/j.devcel.2006.05.010
  6. Xuemei L, Jing Y, Bei X, Juan H, Xinling R, Qun L, et al. Retinoic acid improve germ cell differentiation from human embryonic stem cells. Iran J Reprod Med 2013;11:905-12.
  7. Park TS, Galic Z, Conway AE, Lindgren A, van Handel BJ, Magnusson M, et al. Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells. Stem Cells 2009;27:783-95. https://doi.org/10.1002/stem.13
  8. Ghasemzadeh-Hasankolaei M, Eslaminejad MB, Batavani R, Sedighi-Gilani M. Comparison of the efficacy of three concentrations of retinoic acid for transdifferentiation induction in sheep marrow-derived mesenchymal stem cells into male germ cells. Andrologia 2014;46:24-35. https://doi.org/10.1111/and.12037
  9. Hua J, Qiu P, Zhu H, Cao H, Wang F, Li W. Multipotent mesenchymal stem cells (MSCs) from human umbilical cord: potential differentiation of germ cells. Afr J Biochem Res 2011;5:113-23.
  10. Kurkure P, Prasad M, Dhamankar V, Bakshi G. Very small embryonic-like stem cells (VSELs) detected in azoospermic testicular biopsies of adult survivors of childhood cancer. Reprod Biol Endocrinol 2015;13:122. https://doi.org/10.1186/s12958-015-0121-1
  11. Xie L, Lin L, Tang Q, Li W, Huang T, Huo X, et al. Sertoli cell-mediated differentiation of male germ cell-like cells from human umbilical cord Wharton's jelly-derived mesenchymal stem cells in an in vitro co-culture system. Eur J Med Res 2015;20:9. https://doi.org/10.1186/s40001-014-0080-6
  12. Latifpour M, Shakiba Y, Amidi F, Mazaheri Z, Sobhani A. Differentiation of human umbilical cord matrix-derived mesenchymal stem cells into germ-like cells. Avicenna J Med Biotechnol 2014; 6:218-27.
  13. Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, et al. Derivation of male germ cells from bone marrow stem cells. Lab Invest 2006;86:654-63. https://doi.org/10.1038/labinvest.3700429
  14. Aflatoonian B, Ruban L, Jones M, Aflatoonian R, Fazeli A, Moore HD. In vitro post-meiotic germ cell development from human embryonic stem cells. Hum Reprod 2009;24:3150-9. https://doi.org/10.1093/humrep/dep334
  15. Panula S, Medrano JV, Kee K, Bergstrom R, Nguyen HN, Byers B, et al. Human germ cell differentiation from fetal- and adult-derived induced pluripotent stem cells. Hum Mol Genet 2011;20: 752-62. https://doi.org/10.1093/hmg/ddq520
  16. Song HW, Wilkinson MF. In vitro spermatogenesis: a long journey to get tails. Spermatogenesis 2012;2:238-44. https://doi.org/10.4161/spmg.22069
  17. Easley CA 4th, Phillips BT, McGuire MM, Barringer JM, Valli H, Hermann BP, et al. Direct differentiation of human pluripotent stem cells into haploid spermatogenic cells. Cell Rep 2012;2:440-6. https://doi.org/10.1016/j.celrep.2012.07.015
  18. Bowles J, Koopman P. Retinoic acid, meiosis and germ cell fate in mammals. Development 2007;134:3401-11. https://doi.org/10.1242/dev.001107
  19. Pellegrini M, Di Siena S, Claps G, Di Cesare S, Dolci S, Rossi P, et al. Microgravity promotes differentiation and meiotic entry of postnatal mouse male germ cells. PLoS One 2010;5:e9064. https://doi.org/10.1371/journal.pone.0009064
  20. Tesarik J, Guido M, Mendoza C, Greco E. Human spermatogenesis in vitro: respective effects of follicle-stimulating hormone and testosterone on meiosis, spermiogenesis, and Sertoli cell apoptosis. J Clin Endocrinol Metab 1998;83:4467-73. https://doi.org/10.1210/jcem.83.12.5304
  21. Minaee Zanganeh B, Rastegar T, Habibi Roudkenar M, Ragerdi Kashani I, Amidi F, Abolhasani F, et al. Co-culture of spermatogonial stem cells with sertoli cells in the presence of testosterone and FSH improved differentiation via up-regulation of post meiotic genes. Acta Med Iran 2013;51:1-11.
  22. Reda A, Hou M, Landreh L, Kjartansdottir KR, Svechnikov K, Soder O, et al. In vitro spermatogenesis: optimal culture conditions for testicular cell survival, germ cell differentiation, and steroidogenesis in rats. Front Endocrinol (Lausanne) 2014;5:21.
  23. Gohbara A, Katagiri K, Sato T, Kubota Y, Kagechika H, Araki Y, et al. In vitro murine spermatogenesis in an organ culture system. Biol Reprod 2010;83:261-7.
  24. Huleihel M, Nourashrafeddin S, Plant TM. Application of threedimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta). Asian J Androl 2015;17:972-80. https://doi.org/10.4103/1008-682X.154994
  25. Toyooka Y, Tsunekawa N, Akasu R, Noce T. Embryonic stem cells can form germ cells in vitro. Proc Natl Acad Sci U S A 2003;100:11457-62. https://doi.org/10.1073/pnas.1932826100
  26. Abu Elhija M, Lunenfeld E, Schlatt S, Huleihel M. Differentiation of murine male germ cells to spermatozoa in a soft agar culture system. Asian J Androl 2012;14:285-93. https://doi.org/10.1038/aja.2011.112
  27. Stukenborg JB, Schlatt S, Simoni M, Yeung CH, Elhija MA, Luetjens CM, et al. New horizons for in vitro spermatogenesis? An update on novel three-dimensional culture systems as tools for meiotic and post-meiotic differentiation of testicular germ cells. Mol Hum Reprod 2009;15:521-9. https://doi.org/10.1093/molehr/gap052
  28. Mehrabani D, Hassanshahi MA, Tamadon A, Zare S, Keshavarz S, Rahmanifar F, et al. Adipose tissue-derived mesenchymal stem cells repair germinal cells of seminiferous tubules of busulfan-induced azoospermic rats. J Hum Reprod Sci 2015;8:103-10. https://doi.org/10.4103/0974-1208.158618
  29. West FD, Shirazi R, Mardanpour P, Ozcan S, Dinc G, Hodges DH, et al. In vitro-derived gametes from stem cells. Semin Reprod Med 2013;31:33-8. https://doi.org/10.1055/s-0032-1331795
  30. Childs AJ, Saunders PT, Anderson RA. Modelling germ cell development in vitro. Mol Hum Reprod 2008;14:501-11. https://doi.org/10.1093/molehr/gan042
  31. Jia W, Cheng D, Chen S, Lei L, Wang H. Retinoic acid induces myoblasts transdifferentiation into premeiotic Stra8-positive cells. Cell Biol Int 2011;35:365-72. https://doi.org/10.1042/CBI20100610
  32. Ketkar AA, Reddy KV. Expression pattern of OCT-4 and PLZF transcription factors during the early events of spermatogenesis in mice. J Cell Sci Ther 2012;3:120.
  33. Zhou Q, Li Y, Nie R, Friel P, Mitchell D, Evanoff RM, et al. Expression of stimulated by retinoic acid gene 8 (Stra8) and maturation of murine gonocytes and spermatogonia induced by retinoic acid in vitro. Biol Reprod 2008;78:537-45. https://doi.org/10.1095/biolreprod.107.064337
  34. Liu Y, Niu M, Yao C, Hai Y, Yuan Q, Liu Y, et al. Fractionation of human spermatogenic cells using STA-PUT gravity sedimentation and their miRNA profiling. Sci Rep 2015;5:8084. https://doi.org/10.1038/srep08084
  35. Nagano MC. Techniques for culturing spermatogonial stem cells continue to improve. Biol Reprod 2011;84:5-6. https://doi.org/10.1095/biolreprod.110.088864
  36. Griswold MD. The central role of Sertoli cells in spermatogenesis. Semin Cell Dev Biol 1998;9:411-6. https://doi.org/10.1006/scdb.1998.0203

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