BRL-CM在昆白小鼠胚胎干细胞分离培养中的应用

目的:探讨Buffalo大鼠肝细胞条件培养基(BRL-CM)在昆白小鼠胚胎干细胞(ES)分离培养中的应用。方法:用BRL-CM培养生长在STO(一种小鼠成纤维细胞)饲养层上的昆白小鼠的桑椹胚和囊胚,然后由囊胚内细胞团(ICM)得到ES细胞集落,在有饲养层和无饲养层时用BRL-CM培养分离得到的ES集落。结果:桑椹胚在上述培养条件下能发育成囊胚,然后囊胚在饲养层上长出ICM;用0.1%Trypsin-0.016%EDTA离散ICM,接种到饲养层上培养得到ES集落,ES集落能维持早期传代。结论:在分离培养昆白小鼠的ES细胞时,BRL-CM结合STO饲养层可以用来培养桑椹胚和囊胚,桑椹胚可以发育为囊胚并孵出内细胞团;同时,这种培养条件也能维持昆白小鼠ES细胞的增殖和早期传代。

Application of Buffalo Rat Liver Cell Conditioned Medium in Isolation and Culturing of KM Mouse ES Cells

To probe into the application of the Buffalo rat liver cell conditioned medium(BRL-CM)in culturing the KM mouse'embryonic stem(ES)cell,blastula and morula were cultured on STO feeder cells with BRL-CM.Morula could grow into blastula and hatched from zone pellucida,then attached on the feeder cells together with the collected blastula,followed by the growth of inner cell mass(ICM).ICM were separated onto STO feeder cells or without feeders and cultured with BRL-CM;ES colonies were only gained and passaged in the former case.BRL-CM and STO feeder combining together could maintain ES cells'proliferation and in the undifferentiated state.