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- W2298157085 abstract "Several factors influencing isolation and culturing of Kunming mouse embryonic stem(ES) cell were studied,embryonic fibroblast(MEF) inactivated by mitomycinc was used as the culturing system of the feeder layer.The 3.5 d expanded blastocys and the 4.5 d hatched blastocysts from Kunming mouse were collected respectively,and the attachment and the primary colonies formation rates between the expanded and the hatched blastocysts was compared.The best time of dispersing ICM was studied through collecting blastocysts of 3.5 d,isolating and cloning ES cells by immunosurgery and intact embryo culture.To compare ES cells isolating and cloning by different enzyme concentration,the ES cell was passaged with 0.25% trypsin adding 0.04% EDTA,0.125% trypsin adding 0.02% EDTA and 0.25% trypsin adding 1% chicken serum.The results demonstrated that the attachment rates of the hatched blastocyst was higher than that of the expanded blastocysts(P0.05),but the passage rates was opposite(P0.05).There were not significant differences in the primary colonies formation rates(P0.05).The typical clonies could be formed by means of culturing of 2-3 d(immnunosurgury)and 4-5 d(intact embryo)and picked.The primary colonies formation rates of the 0.25% trypsin adding 1% chicken serum and 0.125% trypsin adding 0.02% EDTA was higher than that of 0.25% trypsin adding 0.04% EDTA(P0.05).There were not significant differences in the primary colonies formation rates between the two groups(P0.05).The experimental results show that the mouse ES cells which are pluripotential cells have been identified by colony morphology,AKP staining and in vitro differentiation." @default.
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- W2298157085 date "2010-01-01" @default.
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- W2298157085 title "Optimization of culture conditions for Kunming species mouse embryonic stem cells." @default.
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