牛兔异种体细胞以兔卵母细胞为核受体的牛体细胞克隆研究

随着同种动物克隆后代的不断降生,人们又开始尝试异种动物体细胞核移植。用兔的卵母细胞作为核受体进行牛的体细胞核移植,研究表明,颗粒细胞作为供核体与兔去核卵母细胞的融合率(60.5%,57.9%)稍高于皮肤成纤维细胞的融合率(52.2%,48.4%),但两者间差异并不显著。颗粒细胞与皮肤成纤维细胞均采用饥饿培养和接触抑制培养两种不同的方式进行诱导处理,融合卵在电激活后能够卵裂并发育的情况差异显著(72.6%对51.4%;70.4%对52.2%)。有少数重构胚发育至囊胚,证明在哺乳动物异种重构胚早期发育中,异种细胞核与细胞质间是相容的。

Cloning of Bovine Adult Somatic Cell with Rabbit's Maturated Oocytes as Nuclear Recipient

With the birth of more and more offsprings from the same breed cloning, people began to study the nuclear transfer from adult somatic cells between two different breeds. In this study, we used the rabbit's maturated oocytes as nuclear recipient to reconstruct embryos from the bovine adult somatic cell. As nuclear donors, the cell fusion efficiency of granulosa cells with the rabbit's enucleated oocytes was higher than that of skin fibroblasts (60.5%,57.9% vs 52.2%,48.4%),but the difference between the two cell types was not significant. Both granulosa cells and skin fibroblasts were grown to confluency or in low serum to induce a quiescent state. The cleaved rate and development of reconstructed embryos was significantly higher with "in low serum" culture method than with "confluency" method (72.6% vs 51.4%;70.4% vs 52.2%). Less reconstructed embryos developed to blastocyst. It demonstrated that the early development of the different breeds' reconstructed embryos could occur in vitro in Mammalia, and the cytoplasm of recipient cell could activate the different breeds' nucleus.