Lei Qi-jing;Pan Qin;Ma Ju-hong;Zhou Zhe;Li Guang-peng;Chen Shu-lin;Hua Jin-lian

Inner Mongolia Univ, Minist Educ, Key Lab Mammalian Reprod Biol & Biotechnol, Hohhot 010021, Peoples R China;Northwest A&F Univ, Coll Vet Med, Shaanxi Ctr Stem Cells Engn & Technol, Yangling 712100, Shaanxi, Peoples R China

male germline stem cells;immortalization;bovine;proliferation;cell transplantation;testis

Male germline stem cells (mGSCs) 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 mGSCs in vitro, especially for large animals such as bovine mGSCs. 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, LIN28A, and CD49F, both in the mRNA 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 mGSCs maintain the criteria of mGSCs.