不同培养系统和传代方法对昆明白小鼠原始生殖细胞分离培养的影响

[目的]寻求适合昆明白小鼠EG细胞的培养系统和传代方法。[方法]常规方法从昆明白小鼠原始生殖细胞中分离EG细胞,用不同培养系统(EG细胞基础培养液,RH-CM,EG细胞基础培养液+MEF)及不同传代方法(机械法,酶消化法,连同饲养层消化法),研究对昆明白小鼠EG细胞分离克隆的影响。[结果]以RH-CM作为培养基效果最好,RH-CM能够更好地促进EG细胞贴壁增殖和集落的形成,有效地维持EG细胞未分化状态;酶消化法和连同饲养层消化法均能获得较高的克隆形成率。[结论]RH-CM和连同饲养层消化法可分别作为昆明白小鼠EG细胞的培养系统和传代方法。

The Effect of Different Culture System and Different Passage Methods on the Isolation and Culture of Embryonic Germ Cells from Kunming White Mouse

[Objective] This experiment is to explore effective method on the isolation and culture of embryonic germ cells from kun ming white mouse.[Mothod]In this study,use different culture medium(EG culture medium,rat heart conditioned medium,EG culture medium with mouse embryonic fibroblast as feeder layer) and different passage methods(passage by hand,direct digestion,digestion together with fibroblast) on the isolation and culture of embryonic germ cells from kun ming white mouse were studied.[Result]Our results showed rat heart conditioned medium had better effects on isolation of EG cells,the clones formation rate was significantly increased compared with EG culture medium group,and the undifferentiation state of EG cells was maintained more efficiently.[Conclusion]The passage methods of digestion together with fibroblast and rat heart conditioned medium can be used for EG cells isolation from kun ming mice.