Anatomy and Cell Biology

Korean Association of Anatomists (대한해부학회)
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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
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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.

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References

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