Demecolcine化学辅助去核技术在牛体细胞核移植中的初步研究

本实验目的是研究demecolcine辅助去核的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期卵母细胞来辅助去除牛卵母细胞核，并用去核的卵母细胞做受体，进行核移植的研究。实验结果显示，demecolcine辅助去核后的卵母细胞质膜有明显一个或二个突起，并且突起内都含有卵母细胞染色体组，显示去核效果较好（57.89%～73.3%）。药物处理一小时为最适时间，去核率可达73.3%。对demecolcine辅助去核的卵母细胞的核移植胚胎发育情况显示囊胚率较盲吸法核移植胚胎较好（12.5%VS10.2%），但二者差异不显著(p>0.05)。Demecolcine药物处理后的卵母细胞能够支持核移植胚胎的发育。Demecolcine辅助去核可以在牛体细胞核移植中的到应用。

The Application of Demecolcine Assisted Oocyte Enucleation on the Bovine Somatic Cell Nuclear Transfer

The aim for this research is to investigate the development ability of bovine SCNT embryo which are produced by the technique of demecolcine assisted oocyte enucleation .The chromosomes of bovine MII oocytes were got rid of by the demecolcine treatment . And the cytoplasm that nuclear had been deleted ,was prepared for the nuclear transfer . As the results showed,the surface of oocytes membrane markedly appeared one or two protrudes which contains the chromosomes and the enucleation effect is good（57.89%～73.3%）.The best period of demecolcine treatment is one hour and the efficiency can 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 is not significantly(p>0.05).The bovine oocytes with demecolcine assisted enucleation can support the development of cloning embryo reconstructed . And this method can be used in the bovine SCNT .