Evaluation of the Effect of Follicle Stimulating Hormone (FSH) on Survival and Colonization of Caprine Spermatogonial Stem Cells during in Vitro Culture

Background and Objectives: Spermatogonial stem cells are specific cells that have the ability of self-renewal and differentiation. These cells play an essential role in maintaining spermatogenesis and fertility. In this regard, the present study was performed with the purpose of investigating the effect of different concentrations of follicle stimulating hormone (FSH) on in vitro colony formation of caprine spermatogonial stem cells. Methods: Spermatogonial cells, were isolated from prepubertal goat testis using two-step enzymatic digestion. Then, isolated cells were cultured for 10 days in four groups. In the control group, simple culture of spermatogonial cells was performed in Dulbecco's Modified Eagle Medium (DMEM) containing 1% antibiotics and 5% FBS (Fetal Bovine Serum). In the treatment groups 1, 2, and 3, different concentrations of follicle stimulating hormone (5, 10, and 20 IU/ml), was added to the culture medium, respectively. The culture media were changed every 72 hours. Identification of cells was confirmed by immunocytochemical staining against PGP9.5 antigen. Immediately after isolation, percentage of cells viability, surface area, and number of colonies formed on 4th, 7th and 10th days after the culture, were evaluated using an inverted microscope. Data were analyzed using one way ANOVA test. Results: The findings indicated that viability rate of Spermatogonial stem cells after isolation was 89.4 ± 2.32%. The effect of FSH on the formation of spermatogonial cells colonies was dose dependent. Doses of 5 and 10 IU/ml increased the surface area and number of the spermatogonial cell derived colonies but dose of 20 IU/ml reduced colonies formation (p < 0.05). Conclusion: FSH can provide an appropriate culture medium for the study of spermatogonial cells in vitro.