Abstract: Spermatogonial stem cells (SSCs) are located on the basement membrane of the testicular seminiferous tubules, which can not only renew themselves but also differentiate to produce sperm. The characteristics of SSCs are critical in assisted reproductive technologies. After years of research, the great progress has been made in in vitro proliferation and differentiation of SSCs from rodent SSCs to human SSCs, which depends on the continuous improvement of in vitro culture methods to a certain extent. At present, the in vitro culture of SSCs has developed from two-dimensional culture to three-dimensional culture, such as soft agar three-dimensional culture method, methylcellulose three-dimensional culture method, nanofiber scaffold three-dimensional culture method, and decellularized biological scaffold three-dimensional culture method. The continuous improvement of SSCs in vitro three-dimensional culture method is of great significance for revealing the mechanism of male germ cell generation and treating male infertility. In order to explore the technical methods for improving the in vitro culture efficiency of SSCs and establish the long-term and stable cell lines of SSCs, the research progress of in vitro three-dimensional culture methods of SSCs were reviewed in this paper.

Key words: Spermatogonia；, Stem cells；, Cell culture techniques；, Infertility, male；, Cell proliferation