Establishment and characterization of immortalized bovine male germline stem cell line

摘要

cqvip:Male germline stem cells(m GSCs) are unique adult germ cells with self-renewal potential and spermatogenesis function in the testis.However,further studies are needed to establish a long-term cultural system of m GSCs in vitro,especially for large animals such as bovine m GSCs.In this study,we first established a stable immortalized bovine male germline stem cell line by transducing Simian virus 40(SV40) large T antigen.The proliferation of these cells was improved significantly.These cells could express spermatogonial stem cell(SSC)-specific markers,such as PLZF,PGP9.5,VASA,LIN28 A,and CD49 F,both in the m RNA and protein levels.Additionally,these cells could be differentiated into three germ layer cells to enter meiosis,form colonies,and proliferate in the seminiferous tubules of busulfan-induced infertile mice.The immortalized bovine m GSCs maintain the criteria of m GSCs.