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。
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.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2012年第12期32-38,F0002,共8页
Journal of Northeast Agricultural University
基金
国际科技合作项目子课题(2011DFA30760-2-1)
东北农业大学博士启动基金(2012RCB27)
关键词
新生牛
雄性生殖干细胞
分离纯化
体外培养
neonatal bull
male germline stem cell
isolation and purification
in vitro culture
