Journal of Integrative Agriculture ›› 2017, Vol. 16 ›› Issue (11): 2547-2557.DOI: 10.1016/S2095-3119(16)61625-8

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  • 收稿日期:2017-02-22 出版日期:2017-11-20 发布日期:2017-11-03

Establishment and characterization of immortalized bovine male germline stem cell line

LEI Qi-jing1, PAN Qin1, MA Ju-hong1, ZHOU Zhe1, LI Guang-peng2, CHEN Shu-lin1, HUA Jin-lian1   

  1. 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering and Technology, Northwest A&F University, Yangling 712100, P.R.China
    2 Key Laboratory for Mammalian Reproductive Biology and Biotechnology, Ministry of Education/Inner Mongolia University, Hohhot 010021, P.R.China
  • Received:2017-02-22 Online:2017-11-20 Published:2017-11-03
  • Contact: Correspondence Hua Jin-lian, E-mail: jinlianhua@nwsuaf.edu.cn; Chen Shu-lin, E-mail: csl_1359@163.com; Li Guang-peng, Tel/Fax: +86-471-4992495, E-mail: gpengli@immu.edu.cn
  • About author:Lei Qi-jing, E-mail: qijinglei@nwafu.edu.cn
  • Supported by:

    This work was supported by the National Major Project for Production of Transgenic Breeding of China (2014ZX08007-002), the National Basic Research Program of China (2016YFA0100203) and the Program of National Natural Science Foundation of China (31572399, 31272518).

Abstract:    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.  

Key words: male germline stem cells ,  immortalization ,  bovine ,  proliferation ,  cell transplantation ,  testis