摘要
探讨了小鼠-牛异质胚构建的简便方法及小鼠体细胞核在牛卵母细胞中重新编程的可能性。以牛的卵母细胞为细胞质供体,用去除透明带及徒手切割的方法去核,设定电压1．5 KV／cm,脉冲时间40μs,与小鼠皮肤成纤维细胞进行电融合的融合率为67．44％,卵裂率为30．23％。融合细胞经离子酶素-6-DMAP激活,用微滴内压制做窝的方法培养小鼠皮肤成纤维细胞异质胚,异质胚的最终发育阶段为8细胞期。结果表明,去透明带牛卵母细胞经切割法去核,可用于小鼠异质胚构建;微滴内做窝的体外培养方法可避免无透明带胚胎的聚合。
Simplified interspecies nuclear transfer in mouse and bovine and the possibilities of the embryos reprogramming were developed. The zona and nucleus of the bovine oocyte were removed by pronase and bisection method respectively. When the skin fibroblast of mouse was fused with the nucleus-free oocyte of the bovine using 40μs and 1.5 kv/cm direct current pulse, the rates of fusion and cleavage were 67.44% and 30.23% respectively. Reconstructed heterogenous embryos finally developed to 8-cell stage when they were previously activated by ionomycin and 6-DMAP and subsequently cultured in press-made micro-weU in the drop of the medium. Zona-free bisection method could be used for mouse heterogenous embryos cloning, and the microwell culture method could be used to avoid zona-free embryos aggregation.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第7期74-79,共6页
China Biotechnology
基金
内蒙古自治区高等学校科学研究资助项目(NJ03118)国家自然科学基金资助项目(30460054)
关键词
小鼠
牛
核移植
体细胞
Mouse Bovine Nuclear transfer Somatic cell
