哺乳动物手工克隆技术与研究进展.docVIP

哺乳动物手工克隆技术与研究进展 　　摘 要：哺乳动物体细胞核移植技术（Somatic Cell nuclear transfer，SCNT）近几十年的发展取得了巨大的成就，但由于需要繁琐的技术、昂贵的显微操作仪且核移植效率不理想，阻碍了核移植克隆的规模化发展。近年来科学界运用了新的革新方法―体细胞手工克隆（Handmade Somatic Cell Cloning，HCM），其操作无需显微操作仪，对操作人的技术要求不高，是纯手工核移植技术。该技术不仅简化操作程序，还提高核移植效率，对今后哺乳动物核移植研究有重要的推动作用。该综述将从核移植技术的研究与进展方面，探讨限制手工克隆（HCM）的因子并以此来优化手工克隆技术。 　　关键词：卵母细胞；手工克隆；去透明带；核移植 　　中图分类号 S855.3 文献标识码 A 文章编号 1007-7731（2018）09-0120-04 　　The Handmade Cloning Technology and Research Progress on Mammalian 　　Wang Julong 　　（Department of Biology Engineering，Wuhu Institute of Technology，Wuhu 241003，China） 　　Abstract：Mammalian somatic cell nuclear transfer has achieved great accomplishments through the past few decades development.However，the limitations of industrialization of nuclear transfer are cumbersome technology as well as expensive equipments and unsatisfied with the efficiency of nuclear transfer.In recent years scientists applied a novel somatic cell cloning method，which called handmade somatic cell cloning（HCM）.Zona-free procedure may offer a solution for the latter problem.It is unnecessary micromanipulation for handmade somatic cell cloning and workers don t require many skills.This technology is not only simplified the operation but also improved the efficiency of somatic cell nuclear transfer，which will advance the development and the industrialization of nuclear transfer to improve the efficiency of somatic cell nuclear transfer in future.This review will from the research progresses on HMC to explore the effects of handmade cloning factors and in order to optimize the handmade cloning technology. 　　Key words：Oocyte；Handmade cloning；Zona-free oocytes；Nuclear transfer 　　自Briggs and King（1952）[1]等人手工操作成功地将两栖类美洲豹蛙（leopard frog）卵母细胞去核以来，科研者不断地完善哺乳动物的核移植技术。1997年，英国《自然》期刊报道了Wilmut 等[2]利用绵羊乳腺细胞进行核移植实验，首次获得世界上第一只克隆绵羊Dolly。这一开创性的实验证实哺乳动物体细胞具有全能性，为后续开展的体细胞核移植提供理论基础。在此之后科学家利用哺乳动物体细胞核移植技术先后成功克隆出猪[3]、牛[4]、兔[5]、小鼠[6]、马[7]等多种哺乳动物，其中在2017年我国科学家首次成功完成克隆猴-“中中”和“华华”。然而核移植克隆的后代表现出体弱，生存力不强易致病死亡甚至出现衰老迹象[8-9]等特征，这些