Indirect Co-Culture of Testicular Cells with Bone Marrow Mesenchymal Stem Cells Leads To Male Germ Cells-SpecificGenes Expression

Background: Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cells-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells (BM-MSCs) in an in vitro indirect co-culture system is investigated. Materials and Methods: Mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid (RA), indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cells-specific markers including Dazl, Piwil2 and Stra8 was evaluated by Real-time PCR. Results: Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not changed significantly in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups. Conclusion: Indirect co-culturing of BMMSCs in the presence of testicular cells leads to expression of male germ cells-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and retinoic acid did not show any positive effect on specific genes expression.

کلیدواژه ها:

BM-MSCs ، Co-culture ، Germ cells ، Retinoic acid ، Testis