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当前位置: 首页 > 文章 > 新生牛雄性生殖干细胞的分离纯化与培养 东北农业大学学报 2012 (12) 32-38+2
Position: Home > Articles > Isolation, purification and culture of male germline stem cells from neo natal bovine testis Journal of Northeast Agricultural University 2012 (12) 32-38+2

新生牛雄性生殖干细胞的分离纯化与培养

作  者：
郑鹏;于磊;田亚光;黄贺;张贵学
单  位：
东北农业大学动物科学技术学院
关键词：
新生牛;雄性生殖干细胞;分离纯化;体外培养
摘  要：
为建立新生牛雄性生殖干细胞的体外增殖与分化体系,将新生牛睾丸消化成单细胞悬液,分别进行差速贴壁、不连续密度梯度Percoll分选雄性生殖干细胞和支持细胞,对分选生殖干细胞进行体外培养。结果表明,差速贴壁法能有效分选雄性生殖干细胞与支持细胞,2%血清浓度的培养液即可用于雄性生殖干细胞的体外培养,添加100 ng·mL-1的GDNF能显著促进雄性生殖干细胞的增殖,增殖形成的雄性生殖干细胞集落具有一定多能性。
译  名：
Isolation, purification and culture of male germline stem cells from neo natal bovine testis
作  者：
ZHENG Peng,YU Lei,TIAN Yaguang,HUANG He,ZHANG Guixue(School of Animal Sciences and Technology,Northeast Agricultural University,Harbin 150030,China)
关键词：
neonatal bull;male germline stem cell;isolation and purification;in vitro culture
摘  要：
In order to establish the method of cell proliferation and differentiation of the newborn calf male germline stem cells(mGSCs) in vitro,calf testis were used in the present study.The method of collagenase and typin was sequentially performed to separate single-cell suspensions,and then used differential adhesion method and discontinued density gradient Percoll sorting to isolate the mGSCs and sertoli cells.For the mGSCs obtained were cultured in vitro.The result showed that the differential adhesion method would seperate the mGSCs and sertoli cells effectively;DMEM/F12 medium containing 2% fetal bovine serum could be used to cultrue mGSCs in vitro.In addition,the medium which added 100 ng.mL-1 GDNF could significantly improve the proliferation rate of mGSCs,and the mGSCs still had pluripotency at the some degree.

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