分离和克隆小鼠ES细胞集落的主要影响因素

The Efficient Derivation of Embryonic Stem Cell Colonies in Kunming Mouse

目的 研究不同培养条件分离和克隆小鼠ES细胞集落的效率。方法 以PMEF饲养层、NIH3T3细胞饲养层或培养液中加入LIF为培养条件 ,分离和克隆昆明小鼠ES细胞集落 ,比较其效率。结果 饲养层的培养条件明显优于培养液中加入LIF的培养条件 ;有饲养层的培养条件下 ,桑椹胚的ES细胞集落出现率显著低于囊胚 ;两种饲养层培养囊胚 ,其ES细胞集落的出现率差异无显著性。结论 以PMEF或NIH3T3细胞作饲养层 ,培养昆明小鼠的囊胚 ,适时离散ICM ,是比较理想的分离ES细胞集落的方法。

Objective To investigate the efficiency of isolation and cloning of ES cell colonies in Kunming mouse. Method\ Three culture conditions were adopted,namely, PMEF feeder layer, NIH3T3 feeder layer and LIF without feeder layer. Results\ According to the formation rate of ES cell colonies, the two kinds of feeder layer are better than LIF only, the blastocysts are better origin for ES cells than morulae and there are no differences between the two kinds of feeder layer. Conclusion It is a more efficient way to isolate and clone ES cell colonies from Kunming mouse blastocysts by PMEF or NIH3T3 feeder layer.