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作 者:桑润滋[1] 韩建永[1] 常万存[2] 仲跻峰[2]
机构地区:[1]河北农业大学动物科技学院,河北保定071001 [2]山东农业科学院生物技术中心,山东济南250100
出 处:《中国兽医学报》2002年第2期194-197,共4页Chinese Journal of Veterinary Science
基 金:河北省教委"九五"攻关课题 ( 9810 9)
摘 要:以兔为模型 ,对影响胚胎细胞核移植重组胚发育力的电融合与激活参数、体外培养系统进行了探索性研究。结果表明 ,有利于重组胚发育力的适宜电融合与激活参数为 2 0 0 V/ m m的电场强度、10 0μs的脉冲宽度 ;在不同的体外培养系统中 ,TCM- 199培养液内于兔胎儿成纤维细胞饲养层 (REF feeder)上共同培养的重组胚获得了最高的 2~ 4-细胞发育率和桑椹胚或囊胚的发育率 ,分别为 6 5 .7%和 2 2 .9%;移植 111枚重组胚给 9只假孕母兔 ,其中 2只妊娠 ,1只完成足月妊娠产出 2只克隆仔兔 ,另 1只在妊娠 2 2 d流产 1只克隆死胎。The low potential for development of reconstructed embryos by nuclear transfer is the primary block for its practical applications.In this experiment,the rabbit was used as a model for embryonic nuclear transfer and the factors affecting the potential of reconstructed embryos such as the parameters of electrofusion and activation,the in vitro cultural systems and the in vivo environment of recipient rabbit were studied.The results showed as follows:The highest rate of electrofusion of EOBC and the best development of reconstructed embryos were obtained by the pulse-strength 200 V/mm and the pulse-width 100 μs.The highest developmental rate for 2-4-cell and morula or blastocyst of the reconstructed embryos were obtained by the co-cultured system on REF feeder in TCM-199 and they were 65.7% and 22.9% respectively.Transfer of 111 nuclear transplant embryos to 9 synchronic recipient rabbits,among which 2 were pregnancy,led to the full development of 3 offspring,2 of them were born normal from one pregnant recipient and one was aborted from the other 22 days after embryonic transfer.
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