山羊-绵羊种间体细胞核移植重构胚构建

Goat-Sheep heterogeneous nuclear transplantation

以山羊皮肤成纤维细胞为供体细胞,绵羊去核卵母细胞为受体细胞进行种间体细胞核移植,构建山羊-绵羊重构胚,并对其采用离子霉素联合6-DM AP法和胞质注射绵羊精子提取物法进行激活,比较两种不同激活方法对山羊-绵羊种间体细胞核移植重构胚的激活效果。结果表明,共构建获得山羊-绵羊重构胚238枚,重构胚在体外能够发育至囊胚阶段;离子霉素联合6-DM AP法激活的卵裂率为58.33%(70/120),囊胚发育率为4.28%(3/70);胞质注射绵羊精子提取物法激活的卵裂率为63.55%(75/118),囊胚发育率为6.66%(5/75),两种方法的激活效果差异不显著。

Totally 238 heterogeneous embryos were reconstructed with goat skin fibroblasts as the donor cells and sheep enucleated oocytes as the recipient,and the effects of two activation methods （Ionomycin combined with 6-DMAP or injection of sperm extract） on the development of heterogeneous reconstructed embryos were compared. The results indicated that a total of 238 heterogeneous embryos were reconstructed,and sheep cytoplast supported mitotic cleavage of goat karyoplasts,and was capable of reprogramming the nucleus to achieve blastocyst stage embryos. The reconstructed embryos activated by Ionomycin combined with f-DAMP reached 58. 33%（70/120） oocyte cleavage rate and 4.28%（3/70） blastocysts rate,while activated by sperm extract reached 63. 55%（75/118） oocyte cleavage rate and 6. 66% （5/75） blastocysts development rate.