Sertoli cell condition medium can induce germ like cells from bone marrow derived mesenchymal stem cells

Document Type: Original Article

Authors

Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Science, Tehran, Iran

Abstract

Objective(s): Although many researchers have confirmed induction of germ cells from bone marrow mesenchymal stem cells (BMMSCs), there are no reports that confirm spontaneous differentiation of germ cells from BMMSCs. In this study, we have evaluated the effect of adult Sertoli cell condition medium (SCCM) as a mutative factor in the induction of germ cells from BMMSCs.
Materials and Methods: BMMSCs were collected from the bone marrow of 6-8-week old NMRI mice and their mesenchymal entities were proven using superficial markers (expression of CD44 and CD73 and non-expresion of CD45 and CD11b) by fow cytometry. Their multi-potential entities were proved with differentiation to osteogenic and adipogenic cells for 21 days. Also isolated Sertoli cells were enriched using lectin coated plates and Sertoli cell condition medium (SCCM) was collected. Sertoli cells were identified by immunocytochemistry and Vimentin marker. The cells were then differentiated into germ cells with SCCM for 2 weeks. Finally induced cells were evaluated by RT-PCR and immunocytochemistry.
Results: Differentiation of mesenchymal stem cells to osteoblast and adipocyte showed their multi-potential property. Expression of CD44 and CD73 and non-expression of CD45 and CD11b confirmed mesenchyme cells. Immunocytochemistry and RT-PCR results showed expression of germ cells specific marker (Mvh).
Conclusion: This study confirmed the effect of SCCM as a motivational factor that can used for differentiation of germ cells from BMMSCs.

Keywords


1. Marques-Mari A, Lacham-Kaplan O, Medrano J, Pellicer A, Simón C. Differentiation of germ cells and gametes from stem cells. Hum Reprod Update. 2009; 15:379-390.

2. Shirazi R, Zarnani AH, Soleimani M, Abdolvahabi MA, Nayernia K, Kashani IR. BMP4 can generate primordial germ cells from bone‐marrow‐derived pluripotent stem cells. Cell Biol Int 2012; 36:1185-1193.

3.Valsangkar S, Bodhare T, Bele S, Sai S. An evaluation of the effect of infertility on marital, sexual satisfaction indices and health-related quality of life in women. J Hum Reprod Sci 2011 ;4:80-85.

4. Alison MR, Poulsom R, Forbes S, Wright NA. An introduction to stem cells. J Pathol 2002; 197:419-423.

5. Hübner K, Fuhrmann G, Christenson LK, Kehler J, Reinbold R, De La Fuente R, et al. Derivation of oocytes from mouse embryonic stem cells. Science 2003; 300:1251-1256.

6. 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;100:11457-11462.

7. 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; 13:727-739.

8. Geijsen N, Horoschak M, Kim K, Gribnau J, Eggan K, Daley GQ. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature 2004; 427:148-154.

9. Shirzeily MH, Pasbakhsh P, Amidi F, Mehrannia K, Sobhani A. Comparison of differentiation potential of male mouse adipose tissue and bone marrow derived-mesenchymal stem cells into germ cells. Iran J Reprod Med 2013; 11:965.

10. Wakitani S, Takaoka K, Hattori T, Miyazawa N, Iwanaga T, Takeda S, et al. Embryonic stem cells injected into the mouse knee joint form teratomas and subsequently destroy the joint. Rheumatology 2003; 42:162-165.

11. Brickman JM, Burdon TG. Pluripotency and tumorigenicity. Nat Genet 2002; 32:557-558.

12. Mohamamadi SM, Movahedin M, Koruji SM. A comparison between the colony formation of adult mouse spermatogonial stem cells in co cultures with sertoli and STO (mouse embryonic fibroblast cell line). Cell J 2010; 12:231-240.

13. Ahmed EA, Barten-van Rijbroek AD, Kal HB, Sadri-Ardekani H, Mizrak SC, van Pelt AM, et al. Proliferative activity in vitro and DNA repair indicate that adult mouse and human Sertoli cells are not terminally differentiated, quiescent cells. Biol Reprod 2009; 80:1084-1091.

14. Lee DR, Kim KS, Yang YH, Oh HS, Lee SH, Chung TG, et al. Isolation of male germ stem cell-like cells from testicular tissue of non-obstructive azoospermic patients and differentiation into haploid male germ cells in vitro. Hum Reprod 2006; 21:471-476.

15. Sousa M, Cremades N, Alves C, Silva J, Barros A. Developmental potential of human spermatogenic cells co-cultured with Sertoli cells. Hum Reprod 2002; 17:161-172.

16. Shamekh R, Saporta S, Cameron DF, Willing AE, Sanberg CD, Johe K, et al. Effects of Sertoli cell-conditioned medium on ventral midbrain neural stem cells: a preliminary report. Neurotox Res 2008; 13:241-246.

17. Geens M, Sermon KD, Van de Velde H, Tournaye H. Sertoli cell-conditioned medium induces germ cell differentiation in human embryonic stem cells. J Assist Reprod Genet 2011; 28:471-480.

18. Huang P, Lin LM, Wu XY, Tang QL, Feng XY, Lin GY, et al. Differentiation of human umbilical cord Wharton's jelly‐derived mesenchymal stem cells into germ‐like cells in vitro. J Cell Biochem 2010; 109:747-754.

19. Lacham‐Kaplan O, Chy H, Trounson A. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. Stem Cells 2006; 24:266-273.

20. Rastegar T, Minaee MB, Roudkenar MH, Kashani IR, Amidi F, Abolhasani F, et al. Improvement of expression of α6 and β1 Integrins by the co-culture of adult mouse spermatogonial stem cells with SIM mouse embryonic fibroblast cells (STO) and growth factors. Iran J Basic Med Sci 2013; 16:134.

21. Scarpino S, Morena AR, Petersen C, Fröysa B, Söder O, Boitani C. A rapid method of Sertoli cell isolation by DSA lectin, allowing mitotic analyses. Mol Cell Endocrinol 1998; 146:121-127.

22. Anway MD, Folmer J, Wright WW, Zirkin BR. Isolation of Sertoli cells from adult rat testes: an approach to ex vivo studies of Sertoli cell function. Biol Reprod 2003; 68:996-1002.

23. Baazm M, Abolhassani F, Abbasi M, Habibi Roudkenar M, Amidi F, Beyer C. An improved protocol for isolation and culturing of mouse spermatogonial stem cells. Cell Reprogram 2013; 15:329-336.

24. Hou J, Yang S, Yang H, Liu Y, Liu Y, Hai Y, et al. Generation of male differentiated germ cells from various types of stem cells. Reproduction 2014; 147:R179-R188.

25. Hua J, Pan S, Yang C, Dong W, Dou Z, Sidhu KS. Derivation of male germ cell-like lineage from human fetal bone marrow stem cells. Reprod Biomed Online 2009; 19:99-105.

26. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006; 8:315-317.

27. 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; 86:654-663.

28. Ghasemzadeh-Hasankolai M, Batavani R,
Eslaminejad MB, Sedighi-Gilani M. Effect of zinc ions on differentiation of bone marrow-derived mesenchymal stem cells to male germ cells and some germ cell-specific gene expression in rams. Biol Trace Elem Res 2012; 150:137-146.

29. Drusenheimer N, Wulf G, Nolte J, Lee JH, Dev A, Dressel R, et al. Putative human male germ cells from bone marrow stem cells. Soc Reprod Fertil Suppl 2006; 63:69-76.

30. Oatley JM, Brinster RL. The germline stem cell niche unit in mammalian testes. Physiol Rev 2012; 92:577-595.

31. Zanganeh BM, Rastegar T, Roudkenar MH, Kashani IR, Amidi F, Barbarestani M. 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.

32. West FD, Mumaw JL, Gallegos-Cardenas A, Young A, Stice SL. Human haploid cells differentiated from meiotic competent clonal germ cell lines that originated from embryonic stem cells. Stem Cells Dev 2010; 20:1079-1088.

33. Yue F, Cui L, Johkura K, Ogiwara N, Sasaki K. Induction of mid brain dopaminergic  neurons from primate embryonic stem cells by coculture with Sertoli cells. Stem cells 2006; 24: 1695-1706.

34. Hai Y, Hou J, Liu Y, Liu Y, Yang H, Li Z, et al. editors. The roles and regulation of Sertoli cells in fate determinations of spermatogonial stem cells and spermatogenesis. Semin Cell Dev Biol 2014; 29:66-75.