电融合/激活对于巴马小型猪体细胞克隆胚胎生产的影响

The effect of electric fusion and activation on the production of Bama mini-pig(Sus scrofa) cloned embryos

本试验旨在建立适用于猪体细胞克隆的融合/激活体系(试验1),优化融合/激活条件以提高重构卵融合率和激活率,为生产足够数量的克隆胚胎用于胚胎移植最终诞生体细胞克隆猪奠定条件。首先比较脉冲时间确定适合体细胞克隆的最佳融合/激活参数,然后利用此参数验证巴马小型猪耳部成纤维细胞克隆胚胎的发育潜能。结果显示:场强100 V/mm,3次电脉冲,脉冲时间为30μs时适于猪体细胞克隆。在试验2中,利用巴马小型猪耳部成纤维细胞为供体细胞构建体细胞克隆胚胎,得到了较高的卵裂率(83.1%)和囊胚发育率(12.4%)。

To obtain enough SCNT embryos for embryos transfer,the study established and optimized the fusion and activation method for SCNT embryos.First,the experiment established the optimal pulse time to the SCNT embryos;second,the experiment investigated the development capacity of the SCNT embryos cloned from Bama mini-pig ear fibroblast cells.The results showed that in experiment 1,pulse number 30 μs group had higher fused rate（70.4 % vs 59.1%,P0.05）and blastocyst rate（10% vs 6.3%,P0.05） than 10 μs group;higher cleaved rate（84.4% vs 66.1%,P0.05） and blastocyst rate（7.4% vs 4.6%,P0.05） than 50 μs group.And in experiment 2,the Bama mini-pig ear fibroblast cell cloned embryos obtained a high cleaved rate（83.1%）and blastocyst rate（12.4%）.It is concluded that in our laboratory,an available method of electric fusion and activation is established to cloned Bama mini-pig SCNT embryos.