Isolation, culture and differentiation of human spermatogonial stem cells into differentiated germ cells, osteoblasts and adipocytes

Spermatogonial Stem Cells (SSCs) differentiate into adult germ cells during spermatogenesis and cause male fertility. Maintenance of SSCs and induction of spermiogenesis in vitro may provide a therapeutic strategy to treat male infertility. Moreover, it is possible to use the SSCs for research in the field of biotechnology and regenerative medicine through differentiation of them into other cell lines.

SSCs were isolated enzymatically from digested testicular tissue and then were cultured. The identity of SSCs were confirmed by anti-OCT4, anti-PLZF through immunocytochemistry. SSCs were cultured in differentiated culture medium. Then, expression level of SCP3, Boule, Crem, Protamine 2 genes and also expression level of SCP3, Protamine2 proteins was determined by Real-time PCR and immunocytochemistry, respectively. In addition, the potency of SSCs differentiation into osteoblasts and adipocytes was evaluated by Alizarin Red and Oil Red staining, respectively.

The results showed that SSCs can differentiate into adult germ cells in in vitro condition. Also, under differentiation conditions, these cells can be distinguished from other cell lines.

The results of this study showed that, given the differentiation potency of SSCs, these cells can be used for the treatment of male infertility, as well as cell therapy and regenerative medicine.