【摘要】 通过比较LIF和大鼠心肌条件液在小鼠ES细胞分离培养过程中的差异,从而选择较为合适的培养液用于小鼠ES细胞的分离培养与深入研究。取怀孕3.5d小鼠囊胚,培养于小鼠胎儿成纤维细胞饲养层上,然后根据ES细胞培养液的不同分成2组,一组添加LIF的ES细胞培养液,另一组添加由大鼠心肌条件液组成的ES细胞培养液。结果显示,小鼠ICM的孵出率在心肌条件液中为76.30%,LIF条件液中为59.35%,两者差异极显著(p<0.01);在LIF条件液中比心肌条件液中能较早地分离出ICM,时间差分别为11、11、12、10和12h,平均为11.2h;对传代的ES细胞集落,培养48h时周边出现分化现象的ES细胞集落所占的比例在心肌条件液中为51.55%,LIF条件液中为31.69%,两者差异显著(p<0.05);第五代小鼠ES细胞核型正常率在心肌细胞条件液中为78.6%,稍高于LIF条件液中的76%,差异不显著。更多还原

【Abstract】 By comparing the difference between LIF media conditioned and rat cardiomyocyte media conditioned during isolation and culture of mouse ES cell, suitable media is selected in the future. Mouse embryos of 3.5 d were collected, cocultured with mouse fetal fibroblasts as feeder layer. According to the difference of ES cell media, mouse embryos were divided into two groups, one with LIF media, other with cardiomyocyte media conditioned. Results showed that isolation rate of ICM with cardiomyocyte media conditioned was 76.30 %, LIF media 59.35 %, and difference between two media was significant (p<0.01). Isolation of mouse ICM with LIF media was earlier than that with cardiomyocyte media conditioned, and difference between two medias were 11, 11, 12, 10 and 12 h respectively, mean difference was 11.2 h. Differentiation rate of ES cell masses of 48th h with cardiomyocyte media conditioned was 51.55 %, LIF media was 31.69 %, and difference between two medias was significant (p<0.05). In addition, Normal karyotype rate of mouse ES cell with cardiomyocyte media conditioned 78.6 % was slightly higher than that with LIF media 76 %, but difference was not significant.更多还原