Demecolcine化学辅助去核技术在牛体细胞核移植中的初步研究  

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

作　　者：沙伟[1] 郭振华[2] 刘赫[1] 殷志明[1] 马红[2] 李仲秋[2] 吴赛辉[2] 刘娣 

机构地区：[1]黑龙江省哈尔滨东北农业大学动物科技学,院哈尔滨150030 [2]黑龙江省农业科学院畜牧研究所胚胎工程实验室,哈尔滨150086

出　　处：《中国生物工程杂志》2009年第2期87-91,共5页China Biotechnology

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。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 1 h and the enucleation efficiency could reach 73. 3%. The blastocyst development of embryo reconstructed with demeeolcine 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 .

关 键 词：秋水仙素 化学辅助去核 整细胞注入 体细胞核移植 

分 类 号：Q819[生物学—生物工程]