大鼠精原干细胞的高效分离和纯化方法

Objective: Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis, are the cloning of a permanent source of differentiation into sperm, it can either self-renew to maintain the number of stem cells in vivo, or differentiate into various stages of spermatogonia until the mature sperm. Because of the samll number of SSCs in the testis, it is very difficult to isolate the SSCs from the testes. This article aims to introduce a highly efficient separation and purification of rat spermatogonial stem cells methods. Methods: In this paper, we used 22-25-day-old Wistar-Iamichi rats. First, we obtained testicular cell suspension by two-step enzymatic digestion, then, according to the different adherence capabilities of SSCs and somatic cells (mainly setoli cells) and other differentiated spermatogonia, we purified the rat spermatogonial stem cells. Results: We obtained 3×105 stem cells from five male rats. The SSCs can form clones during in vitro culture, and the clonings can express specific marker genes of SSCs, such as GFRα1 and CDH1. Conclusion: this paper describes the efficient method of separation and purification of rat SSCs, this method is simple and the purified SSCs are with high viability and proliferating capacity. The method also opened up a new road for the isolation and purification of large animal and human SSCs in the near future.