Abstract: Recently, spermatogonial stem cell (SSC), the postnatal male germ-line stem cells, has been a subject of intense and increasing interest as a result of exciting developments in stem cell research in general and the development of new research tools allowing the isolation, culture and transplantation of these unique cells. SSC is a adult stem cell population resided in the basal compartment of the seminiferous epithelium, which is not only capable of self-renewal and differentiation, but the only adult stem cells in the normal postnatal body that undergo self-renewal throughout life and transmit gene to the progeny. In order to obtain stem cells enough for researches and application, understanding the regulation mechanisms of the proliferation and differentiation of stem cells is the common problem in stem cell research. With the aid of SSCs transplantation, a functional assay for SSCs, the culture conditions of SSCs have been examined deeply in recently years. This review focuses on what is know about the maintenance of SSCs in vitro and discusses the possible problem in future studies.