小型猪克隆胚的构建及胚胎移植研究

目的建立小型猪的体细胞核移植技术，为开展转基因小型猪动物模型研究提供技术支撑。方法应用体细胞核移植技术，以小型猪胎儿成纤维细胞和枫泾猪耳成纤维细胞作为供体细胞进行体细胞核移植，并将两种重构胚混合后进行胚胎移植。结果两种供体细胞在体外发育能力上无显著差异（P〉0．05）；电融合后进行化学激活能部分提高囊胚发育率，但差异不显著（P〉0．05）。两种方法生产的小型猪和枫泾猪克隆胚胎混合后，经胚胎移植均能顺利产下4只健康后代。经微卫星亲缘关系鉴定，克隆猪lOO％与供体细胞相同，与代孕母猪无关。结论建立的克隆平台能够用于克隆猪的生产。

Studies on Construction and Transfer of Cloned Minipig Embryos

Objective To establish the somatic cell nuclear transfer technology of minipig and provide technical support for studies of transgenic animal models. Methods Using somatic cell nuclear transfer techniques, nuclear transfer was finished with fetal fibroblast cells and ear fibroblast cells （donor cells） from minipig and Fengjing pig respectively. Embryo transfer was carried out using the two kinds of reconstructed embryos. Result There were no significant differences between the two kinds of donor cells in vitro development （P〉0.05）. Chemical activation after electric fusion could partially improve the blastocyst rates, but the difference was not significant （P〉0.05）. Four healthy cloned piglets were borned after embryos transfer by the mixture of the two kinds reconstructed embryos. Microsatellite identifica- tion of cloned piglets showed whole same with their donor cells in heredity, and had nothing to do with their surrogate sows. Conclusion The method of our study group could be used to produce clone pigs.