供体细胞状态对牛重组胚发育的影响李向臣，跃华，刘鹏，乌云其其格，关伟军，马月辉**（中国农业科学院北京畜牧兽医研究所，北京100094）摘要：本试验通过比较供体成纤维细胞的不同血清饥饿培养天数、传代次数、冻存复苏等方面试验条件对重组胚发育的影响因素进行了深入的研究。结果表明：供体细胞血清饥饿0、1～3、4～6、7～9d之间重组胚卵裂率没有显著差异（P＞0.05），但囊胚率以饥饿1～3d的最高，与4～6和7～9d组别差异显著（P＜0.05）；以传代0、1～3、4～6、7～9代的细胞作为供体细胞，卵裂率没有显著差异（P＞0.05），桑椹胚率以传1～3代最高，差异显著（P＜0.05）；2代细胞解冻后的卵裂率显著高于4和8代细胞，而以2和6代冷冻解冻后细胞为供体，卵裂率并无明显差异，8代细胞的桑椹胚率显著低于其它组。本试验为提高供体成纤维细胞在卵母细胞中重新程序化及后续重组胚发育等相关研究提供参考。关键词：牛；供体成纤维细胞；血清饥饿；核移植中图分类号：Q322 文献标识码：A 文章编号：0366-6964（2008）04-0513-04The Research of the Impact of the State of Donor Cell on the Growth of Reorganization EmbryosLI Xiang-chen，YUE Hua，LIU Peng,WU Yun Qi Qi Ge,GUAN Wei-jun,MA Yue-hui**(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100094,China)Abstract:This study conducted in-depth research on the effect factors of the growth of recon-structed embryos,through comparing the different serum starvation days,the number of genera-tions with the recovery of cryopreserved of Donor fibroblast and So text conditions.The research results show that the difference of reorganized embryo cleavage rate is not significant（P＞0.05）as donor serum were starved 0,1-3,4-6 and 7-9 days,but blastocysts rate was the highest as do-nor serum were starved 1-3 days,and the difference was significant（P＜0.05）compared with the4-6 and 7-9 days groups;the difference of embryo cleavage rate wasnot significant（P＞0.05）and morula rate of 1—3 generation was the highest as the cells of 0、1-3、4-6 and 7-9 generation as a source of donor cells;The cleavage rate of reorganized embryos from 2 generation frozen-thawed cell was significantly higher compared with 4 and 8 generation.2 and 6 generation thawed cells of after frozen as were donor cells,egg(embryo)cleavage rate did not differ significantly,and mor-ula rate of 8 generation was lower compared with the other groups.This experiment provide im-portant theoretical basis and technological support to improve donor cells re-program(procedure)in oocyte,the growth of follow-up reorganization embryo and the research of related fields.Key words:bovine;donor fibroblast;serum starvation;nuclear transfer自从高度分化的体细胞在卵母细胞质中具有去分化、重获全能性的能力被证实之后，体细胞核移植收稿日期：2006-11-13的研究得到了飞速的发展，已有很多种类的体细胞被应用为核供体，并成功获得了体细胞核移植的小基金项目：国家自然科技资源平台建设项目（2004DKA30450）；国家“863”高技术项目（2006AA102198）作者简介：李向臣（1973-），男，内蒙古通辽人，博士，主要从事哺乳动物胚胎工程和发育生物学研究，E-mail:nobelli@126.com*通讯作者：关伟军，博士，教授，博导，主要从事细胞与分子生物学研究，E-mail:wjguan86@126.com马月辉，男，研究员，博导，E-mail:yue-hui.ma@263.net
供体细胞状态对牛重组胚发育的影响
