一种有效分离及在体外扩增成年小鼠精原干细胞的方法  

Effective Isolation and Propagation of Adult Mouse Spermatogonial Stem Cells in vitro

作　　者：冉竞超[1,2] 曹文广[2] 孙红艳[2,3] 王令[1] 辛颖[1] 张智英[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]中国农业科学院北京畜牧兽医研究所,北京100193 [3]南京农业大学动物科技学院,南京210014

出　　处：《西北农业学报》2012年第10期1-6,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：农业部转基因重大专项(2011ZX08008-003)

摘　　要：尝试建立一种简便有效的成年小鼠精原干细胞分离培养及扩增的方法。从单只成年小鼠的睾丸中经过两步酶消化法分离精原干细胞,并在去除间质细胞、纯化后的支持细胞饲养层上培养与扩增,获得稳定增殖的精原干细胞系。此方法可显著提高建系成功率,保证细胞系的单一基因型,显著降低体外培养精原干细胞的成本。为成体基因来源的转基因动物的制作和疾病模型的建立提供技术支持。In our study,we established a convenient and effective method for isolating and propagating the adult spermatogonial stem cells in vitro.We isolated the spermatogonial stem cells from the testes of one adult KM mouse by two-step digestion,and then cultured the cells on the purified Sertoli cells treated with mitomycin-C,eventually we established the stable adult spermatogonial stem cell lines.Our study increased the efficiency of cell line establishment prominently,guaranteed to achieve the single genotype cell line,and reduced the cost of culturing adult male germline stem cells in vitro.This method provided the foundation for producing the transgenic animals and disease models derived from adult genotype.

关 键 词：成年小鼠 精原干细胞 支持细胞 间质细胞 

分 类 号：Q813.11[生物学—生物工程] Q492.4