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KMID : 1237720230560040508
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Anatomy & Cell Biology
2023 Volume.56 No. 4 p.508 ~ p.517
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Evaluating the effect of conditioned medium from mesenchymal stem cells on differentiation of rat spermatogonial stem cells
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Hoda Fazaeli
Mohsen Sheykhhasan
Naser Kalhor
Faezeh Davoodi Asl
Mojdeh Hosseinpoor Kashani
Azar Sheikholeslami
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Abstract
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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 mesenchymalstem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this studywas 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 WistarRats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco’smodified 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 levelof differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL andDDX4 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 DAZLand DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL andDDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. Inthis study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to inducethe differentiation in SSCs.
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KEYWORD
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Spermatogonial stem cells, Mesenchymal stem cells, Conditioned medium, Differentiation
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