不同方法处理供核细胞对细胞周期和重构胚发育的影响

摘要

To compare the cell relative proportion at G0/G1 phase, chromosome karyotypes, and efficiency of bovine somatic cell nuclear transfer between the transgenic donor cells and non-transgenic donor cells synchronized by serum starvation or contact inhibition. The proportions of the donor cells at the G0/G1 phase were counted by fluorescence activated cell sorting (FACS) and cell chromosome karyotypes were assessed at the same time course. It was found that serum starvation or contact inhibition could effectively synchronize at G0/G1 phase in both transgenic and non-transgenic donor cells, and almost no difference in the two groups. However, there was a significant difference between the both group and untreated control (P＜0.05). The serum starvation resulted in better efficiency of nuclear transfer than the inhibition group and control(P＜0.05). Moreover, when the donor cells were cultured to the 6th day, a higher percentage of abnormal karyotypes were observed as compared to the 2rd day or 4th day. Taken together, the donor cells treated for less than 4 days was optimal for cell cycle synchronization and nuclear transfer.%利用流式细胞仪和细胞染色体核型分析技术,比较奶牛的转基因体细胞和正常细胞经血清饥饿、抑制培养周期同步化处理后的G0/G1期细胞比例；并将同步化处理的核供体细胞进行核移植,然后统计囊胚发育率.结果表明,血清饥饿和抑制培养均能获得较高比例的G0/G1期细胞,两组间差异不显著(P＞0.05),但均显著高于未处理对照组(P＜0.05)；血清饥饿组的囊胚率显著高于抑制培养组和非处理对照组(P＜0.05)；但细胞同步化处理6d后细胞染色体核型异常率增加.因此,要获得正常核型的G0/G1核移植供体细胞和较高的囊胚率,同步化处理时间以不超过4d为宜.