Anatomy and Cell Biology

Korean Association of Anatomists (대한해부학회)
권호 표지
DOI QR코드

DOI QR Code

Evaluating the effect of conditioned medium from mesenchymal stem cells on differentiation of rat spermatogonial stem cells

  • Hoda Fazaeli (Department of Mesenchymal Stem Cells, Academic Center for Education, Culture, and Research (ACECR)) ;
  • Mohsen Sheykhhasan (Department of Mesenchymal Stem Cells, Academic Center for Education, Culture, and Research (ACECR)) ;
  • Naser Kalhor (Department of Mesenchymal Stem Cells, Academic Center for Education, Culture, and Research (ACECR)) ;
  • Faezeh Davoodi Asl (Department of Mesenchymal Stem Cells, Academic Center for Education, Culture, and Research (ACECR)) ;
  • Mojdeh Hosseinpoor Kashani (Department of Biology, Faculty of Sciences, Central Tehran Branch, Islamic Azad University) ;
  • Azar Sheikholeslami (Department of Mesenchymal Stem Cells, Academic Center for Education, Culture, and Research (ACECR))
  • Received : 2022.11.30
  • Accepted : 2023.08.21
  • Published : 2023.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

In cancer patients, chemo/radio therapy may cause infertility by damaging the spermatogenesis affecting the self-renewal and differentiation of spermatogonial stem cells (SSCs). In vitro differentiation of stem cells especially mesenchymal stem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this study was to evaluate the proliferation and differentiation of SSCs using their co-culture with Sertoli cells and conditioned medium (CM) from adipose tissue-derived MSCs (AD-MSCs). Testicular tissues were separated from 2-7 days old neonate Wistar Rats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1X antibiotic, basic fibroblast growth factor, and glial cell line-derived neurotrophic factor. The cells were treated with the CM from AD-MSCs for 12 days and then the expression level of differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL and DDX4 was calculated. Scp3, Dazl, and Prm1 were significantly increased after treatment compared to the control group, whereas no significant difference was observed in Stra8 expression. The immunocytochemistry images showed that DAZL and DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL and DDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. In this study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to induce the differentiation in SSCs.

Keywords

References

  1. Matzuk MM, Lamb DJ. The biology of infertility: research advances and clinical challenges. Nat Med 2008;14:1197-213. 
  2. Vander Borght M, Wyns C. Fertility and infertility: Definition and epidemiology. Clin Biochem 2018;62:2-10. 
  3. Cooper TG, Noonan E, von Eckardstein S, Auger J, Baker HW, Behre HM, Haugen TB, Kruger T, Wang C, Mbizvo MT, Vogelsong KM. World Health Organization reference values for human semen characteristics. Hum Reprod Update 2010;16:231-45. 
  4. Babakhanzadeh E, Nazari M, Ghasemifar S, Khodadadian A. Some of the factors involved in male infertility: a prospective review. Int J Gen Med 2020;13:29-41. 
  5. Kostereva N, Hofmann MC. Regulation of the spermatogonial stem cell niche. Reprod Domest Anim 2008;43(Suppl 2):386-92. 
  6. Vahdati A, Fathi A, Hajihoseini M, Aliborzi G, Hosseini E. The regenerative effect of bone marrow-derived stem cells in spermatogenesis of infertile hamster. World J Plast Surg 2017;6:18-25. 
  7. Goharbakhsh L, Mohazzab A, Salehkhou S, Heidari M, Zarnani AH, Parivar K, Akhondi MM. Isolation and culture of human spermatogonial stem cells derived from testis biopsy. Avicenna J Med Biotechnol 2013;5:54-61. 
  8. Meistrich ML. Effects of chemotherapy and radiotherapy on spermatogenesis in humans. Fertil Steril 2013;100:1180-6. 
  9. Anderson RA, Mitchell RT, Kelsey TW, Spears N, Telfer EE, Wallace WH. Cancer treatment and gonadal function: experimental and established strategies for fertility preservation in children and young adults. Lancet Diabetes Endocrinol 2015;3:556-67. 
  10. Marettova E, Maretta M, Legath J. Toxic effects of cadmium on testis of birds and mammals: a review. Anim Reprod Sci 2015;155:1-10. 
  11. Trondle I, Westernstroer B, Wistuba J, Terwort N, Schlatt S, Neuhaus N. Irradiation affects germ and somatic cells in prepubertal monkey testis xenografts. Mol Hum Reprod 2017;23:141-54. 
  12. Park KS, Bandeira E, Shelke GV, Lasser C, Lotvall J. Enhancement of therapeutic potential of mesenchymal stem cell-derived extracellular vesicles. Stem Cell Res Ther 2019;10:288. 
  13. Almeria C, Weiss R, Roy M, Tripisciano C, Kasper C, Weber V, Egger D. Hypoxia conditioned mesenchymal stem cell-derived extracellular vesicles induce increased vascular tube formation in vitro. Front Bioeng Biotechnol 2019;7:292. 
  14. Bazoobandi S, Tanideh N, Rahmanifar F, Zare S, Koohi-Hosseinabadi O, Razeghian-Jahromi I, Dianatpour M, Ahmadi M, Khoradmehr A, Nabipour I, Khodabandeh Z, Tamadon A. Preventive effects of intrauterine injection of bone marrow-derived mesenchymal stromal cell-conditioned media on uterine fibrosis immediately after endometrial curettage in rabbit. Stem Cells Int 2020;2020:8849537. 
  15. Nagano M, Ryu BY, Brinster CJ, Avarbock MR, Brinster RL. Maintenance of mouse male germ line stem cells in vitro. Biol Reprod 2003;68:2207-14. 
  16. Zhang D, Liu X, Peng J, He D, Lin T, Zhu J, Li X, Zhang Y, Wei G. Potential spermatogenesis recovery with bone marrow mesenchymal stem cells in an azoospermic rat model. Int J Mol Sci 2014;15:13151-65. 
  17. Vlajkovic S, Cukuranovic R, Bjelakovic MD, Stefanovic V. Possible therapeutic use of spermatogonial stem cells in the treatment of male infertility: a brief overview. ScientificWorldJournal 2012;2012:374151. 
  18. Forbes CM, Flannigan R, Schlegel PN. Spermatogonial stem cell transplantation and male infertility: current status and future directions. Arab J Urol 2017;16:171-80. 
  19. Phillips BT, Gassei K, Orwig KE. Spermatogonial stem cell regulation and spermatogenesis. Philos Trans R Soc Lond B Biol Sci 2010;365:1663-78. 
  20. Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, Gromoll J, Engel W. Derivation of male germ cells from bone marrow stem cells. Lab Invest 2006;86:654-63. 
  21. Ghasemzadeh-Hasankolaei M, Eslaminejad MB, Sedighi-Gilani M. Derivation of male germ cells from ram bone marrow mesenchymal stem cells by three different methods and evaluation of their fate after transplantation into the testis. In Vitro Cell Dev Biol Anim 2016;52:49-61. 
  22. Yu Z, Ji P, Cao J, Zhu S, Li Y, Zheng L, Chen X, Feng L. Dazl promotes germ cell differentiation from embryonic stem cells. J Mol Cell Biol 2009;1:93-103. Erratum in: J Mol Cell Biol 2010;2:170. 
  23. Li H, Liang Z, Yang J, Wang D, Wang H, Zhu M, Geng B, Xu EY. DAZL is a master translational regulator of murine spermatogenesis. Natl Sci Rev 2019;6:455-68. 
  24. Barkholt L, Flory E, Jekerle V, Lucas-Samuel S, Ahnert P, Bisset L, Buscher D, Fibbe W, Foussat A, Kwa M, Lantz O, Maciulaitis R, Palomaki T, Schneider CK, Sensebe L, Tachdjian G, Tarte K, Tosca L, Salmikangas P. Risk of tumorigenicity in mesenchymal stromal cell-based therapies--bridging scientific observations and regulatory viewpoints. Cytotherapy 2013;15:753-9. 
  25. Maumus M, Jorgensen C, Noel D. Mesenchymal stem cells in regenerative medicine applied to rheumatic diseases: role of secretome and exosomes. Biochimie 2013;95:2229-34. 
  26. Kadam P, Van Saen D, Goossens E. Can mesenchymal stem cells improve spermatogonial stem cell transplantation efficiency? Andrology 2017;5:2-9. 
  27. Moreno SG, Attali M, Allemand I, Messiaen S, Fouchet P, Coffigny H, Romeo PH, Habert R. TGFbeta signaling in male germ cells regulates gonocyte quiescence and fertility in mice. Dev Biol 2010;342:74-84. 
  28. Minaee Zanganeh B, Rastegar T, Habibi Roudkenar M, Ragerdi Kashani I, Amidi F, Abolhasani 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. 
  29. Spradling A, Drummond-Barbosa D, Kai T. Stem cells find their niche. Nature 2001;414:98-104. 
  30. Scadden DT. The stem-cell niche as an entity of action. Nature 2006;441:1075-9. 
  31. Takashima S, Kanatsu-Shinohara M, Tanaka T, Morimoto H, Inoue K, Ogonuki N, Jijiwa M, Takahashi M, Ogura A, Shinohara T. Functional differences between GDNF-dependent and FGF2-dependent mouse spermatogonial stem cell self-renewal. Stem Cell Reports 2015;4:489-502.