不同联合激活方法对牛体细胞核移植胚胎体外发育的影响  

Influence of Different United Activation on Development of Bovine Somatic Cell Transfer Embryo

作　　者：杨洪武[1] 齐晓龙[1] 熊善辉[1] 倪和民[1] 刘云海[1] 郭勇[1] 杨佐君[1] 

机构地区：[1]北京农学院动物科学技术学院,北京102206

出　　处：《中国农学通报》2015年第11期59-62,共4页Chinese Agricultural Science Bulletin

基　　金：2013年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目"体细胞转基因克隆肉牛新品系培育与利用"(PXM2013_014207_000067);2013年度国家自然基金面上项目"牛体内外植入前胚胎IFNτ的差异表达及对子宫上皮细胞ISG15和Wnt7a表达的体外诱导"(31272526)

摘　　要：为摸索牛体细胞核移植最优的联合激活方法,利用体外成熟22～24 h的去核卵母细胞作为受体细胞质,牛耳部成纤维细胞作为供体细胞,注射到透明带间隙。在电融合参数为电场强度25 N/C,脉冲宽度10μs,2次脉冲的融合条件下,对不同联合化学激活方式进行比较。结果显示在电融合参数不变的条件下,6-DMAP＋CB的激活方式,融合率最高但差异不显著,后期发育的卵裂率（74.1%）和囊胚率（27.8%）也最高。笔者初步得到了较适合牛体细胞核移植的联合激活方法,为今后牛的体细胞核移植胚胎的发育和研究奠定了基础。The aim of the study was to explore the best united activation method of bovine somatic cell nucleartransplantation. Enucleated oocytes matured in vitro for 22-24 hours were taken as the recipient cytoplasts, thefibroblasts from bovine ears were taken as donor cells and were inserted into the perivitelline space of theenucleated oocyte to form the reconstructed couplet. The combined chemical activation methods werecompared under the condition of electrical filed strength 25N/C, pulse width 10 μs, pulse number 2 times. Theresults showed that the best chemical activation was 6- DMAP ＋ CB whose rates of cleavage（74.1%） andblastula（27.8%） were higher than those of other groups. The fusion rate of 6-DMAP＋CB was the highest butwas not significantly different compared with that of other groups. The optimal condition for fusion andactivation of the bovine somatic cell nuclear transfer was obtained preliminarily and would make foundation forsomatic nuclear transplantation in the future.

关 键 词：牛 体外成熟 体细胞核移植 化学激活 胚胎培养 

分 类 号：S814.8[农业科学—畜牧学]