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Amini Mahabadi J, Amini Mahabadi A, Faghihi M, Nikzad H, Abolfazl Aazami Tameh A. Differentiation of Mouse Induced Pluripotent Stem Cells (miPSCs) Into Male Germ Cells (mGCs) By Retinoic Acid and Testosterone. SJMR 2021; 6 (1) :61-69
URL: http://saremjrm.com/article-1-217-en.html

Introduction: Donation of sex-specific cells or production of germ cells through germ cells and preparation of the source of these cells by using embryonic stem cells and induced pluripotent stem cells are considered as methods for treating azoospermia, even in the absence of spermatogonia stem cells. The aim of the present study was to investigate the effect of retinoic acid and testosterone on the differentiation of mouse-induced pluripotent stem cells into male germ cells.
Material and methods: Inactivated mouse embryonic fibroblast cells (MEFs) were used as feeding cells for mouse-induced pluripotent stem cells (miPSCs). These stem cells were transformed into embryoid bodies (EBs). Four groups including control, retinoic acid (RA), testosterone (T), and the combined group of retinoic acid with testosterone (RA+T) were adjusted at intervals of days 0, 4,       and 7. The target genes were Stra8 (Stimulated by Retinoic Acid 8), Ddx4 (DEAD-Box Helicase 4), and AKAP3 (A-Kinase Anchoring Protein 3), and their expression was assessed by the Real-Time PCR technique. Statistical data were analyzed by SPSS software, one-way ANOVA, and Tukey test with a P-value of <0.05 regarded as significant.
Results: The data showed that the expression of Stra8 gene on day 0 in RA group had a significant increase compared to T group (P<0.05) and a non-significant increase compared to the control group. Also, a significant increase was observed in RA+T group compared to the control group, as well as the T group (P<0.001). Ddx4 gene expression was not significantly different for different days and groups. The expression of AKAP3 gene was significantly increased between T and RA group on day 4 (P<0.01), but the expression was decreased between group T and RA+T group (P<0.01). On day 7, the expression of this gene was significantly different between RA+T, control, T, and RA groups (P<0.0001).
Conclusion: Control of differentiation of induced pluripotent stem cells (miPSCs) in mouse is very difficult and in this study, the role of testosterone in vitro, in this differentiation process was carefully investigated. For the testosterone-treated group, it was shown that the combined effect of testosterone with retinoic acid enhances the differentiation process.