【摘要】 研究demecolcine辅助去核后的牛卵母细胞能否支持牛的核移植胚胎的发育。通过化学药物demecolcine处理牛MII期卵母细胞后,借助显微操作去除牛卵母细胞核,并将去核的卵母细胞作为受体,构建重构胚后观察发育情况。实验结果显示,demecolcine处理后牛卵母细胞质膜有明显一个或二个突起,且突起内含有卵母细胞染色体,demecolcine辅助牛卵母细胞去核效果较好(57.89%～73.3%),处理时间以1h为宜,去核率可达73.3%。对demecolcine辅助去核后获得的核移植胚胎发育情况显示,囊胚发育率较盲吸法去核核移植胚胎较好(12.5%vs 10.2%),但二者差异不显著(p>0.05)。Demecolcine辅助处理去核技术能有效去除牛卵母细胞核,且去核后的卵母细胞能够支持核移植胚胎的发育。更多还原

【Abstract】 The aim was to investigate the application of demecolcine assisted oocyte enucleation in bovine SCNT,and the development ability of embryos which are produced by this technique.After 19 h maturation,the chromosomes of bovine MII oocytes were got rid of by the demecolcine treatment.And the cytoplasma that nuclear materials had been deleted,was prepared for the nuclear transfer receptor.As the results showed,with the demecolcine treatment,the surface of bovine oocyte membrane markedly appeared one or two protrudes which contains the oocyte chromosomes,and the enucleation effect was good(57.89%～73.3%).The best duration of demecolcine treatment for enucleation in bovine oocytes was 1h and the enucleation efficiency could reach 73.3%.The blastocyst development of embryo reconstructed with demecolcine assisted enucleation oocytes were better than that reconstructed with blind sucking enucleation.(12.5%VS10.2%),but the difference was not significant(p>0.05).The bovine oocytes cytoplasma enucleated with demecolcine treatment could support the development of reconstructed embryo.And this method can be used in the bovine SCNT.更多还原