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Isolation, Cultivation and Identification of Spermatogonial Stem Cells from Juvenile Buffalo Testes

Isolation, Cultivation and Identification of Spermatogonial Stem Cells from Juvenile Buffalo Testes

Huan Yang1, Tingting Li1, Huimin Zhao1,2, Xiaoyuan Zhang1, Huiyan Xu1, Yangqing Lu1, Xingwei Liang1, Shengsheng Lu1, Xiaogan Yang1*and Kehuan Lu1*

1State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, College of Animal Science and Technology. 
2College of Life Science and Technology, Guangxi University, Nanning 530005, China

*      Corresponding author: xgyang@gxu.edu.cn; khlu@gxu.edu.cn

ABSTRACT

Spermatogonial stem cells (SSCs), a class of primitive germ cells located on the basement of the seminiferous tubules of the testes, are the only kind of adult stem cells that can pass their genetic material to offspring and are also capable of proliferation and differentiation, ensuring the efficiency of passing transgenes from parents to subsequent generations. In this research, to understand the growth characteristics of buffalo SSCs in vitro, methods for their isolation, enrichment, culture and preliminary identification were established. Testes from 3- to 6-month-old buffalos were digested with two-step enzymatic treatment to obtain isolated single cells and then enriched with a differential plating method. Isolated SSCs were cultured on STO feeder cells with SSCs culture medium (SSCM), which is serum-free. SSCs began to proliferate on the second day and quickly formed grape-like clusters that were consistent with the morphological features of SSCs. These cells were identified as positive by immunofluorescence staining. This study successfully isolated, enriched and identified buffalo SSCs and established an effective platform to explore the mechanisms of proliferation and differentiation of buffalo SSCs.
 

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Pakistan Journal of Zoology

April

Vol. 53, Iss. 2, Pages 401-800

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