小型猪克隆胚的构建及胚胎移植研究

doi:10.3969/j.issn.1674-5817.2011.05.006

实验动物与比较医学 ›› 2011, Vol. 31 ›› Issue (5): 333-337.DOI: 10.3969/j.issn.1674-5817.2011.05.006

所属专题：实验动物资源开发与利用

小型猪克隆胚的构建及胚胎移植研究

戴建军^1,2, 李垚³, 顾晓龙^1,2, 张廷宇^1,2, 吴彩凤^1,2, 张树山^1,2, 吴斌^1,2, 陈会兰^1,2, 陈学进³, 张德福^1,2

1.上海市农业科学院畜牧兽医研究所, 上海 201106;
2.上海市农业遗传育种重点实验室动物遗传工程研究室, 上海 201106;
3.上海交通大学医学院实验动物科学部, 上海200025

收稿日期:2011-07-15 出版日期:2011-10-15 发布日期:2011-10-25
作者简介:戴建军(1979-),男,临床兽医学,从事胚胎生物技术的研究,E-mail:blackman0520@126.com
基金资助:
国家转基因生物新品种培育科技重大专项(2008ZX08006-005,2009ZX08006-014B);上海市科技兴农攻关项目(沪农科攻字2005-3-5),上海市科技兴农推广项目(沪农科推字2007-3-7)和上海市科委科技成果转化项目(103919N1800)资助

Studies on Construction and Transfer of Cloned Minipig Embryos

DAI Jian-jun^1,2, LI Yao³, GU Xiao-long^1,2, ZHANG Ting-yu^1,2, WU Cai-feng^1,2, ZHANG Shu-shan^1,2, WU Bin^1,2, CHEN Hui-lan^1,2, CHEN Xue-jin³, ZHANG De-fu^1,2

1. Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China;
2. Shanghai Municipal Key Laboratory of Agri-genetics and Breeding, Shanghai, 201106, China;
3. School of Medicine Department of Laboratory Animal Science, Shanghai Jiao Tong University, Shanghai, 200025, China

Received:2011-07-15 Online:2011-10-15 Published:2011-10-25

摘要/Abstract

摘要： 目的建立小型猪的体细胞核移植技术,为开展转基因小型猪动物模型研究提供技术支撑。方法应用体细胞核移植技术,以小型猪胎儿成纤维细胞和枫泾猪耳成纤维细胞作为供体细胞进行体细胞核移植,并将两种重构胚混合后进行胚胎移植。结果两种供体细胞在体外发育能力上无显著差异(P>0.05); 电融合后进行化学激活能部分提高囊胚发育率,但差异不显著(P>0.05)。两种方法生产的小型猪和枫泾猪克隆胚胎混合后,经胚胎移植均能顺利产下4只健康后代。经微卫星亲缘关系鉴定,克隆猪100%与供体细胞相同,与代孕母猪无关。结论建立的克隆平台能够用于克隆猪的生产。

关键词: 体细胞核移植, 克隆, 小型猪, 化学激活

Abstract: Objective To establish the somatic cell nuclear transfer technology of minipig and provide technical support for studies of transgenic animal models. Methods Using somatic cell nuclear transfer techniques, nuclear transfer was finished with fetal fibroblast cells and ear fibroblast cells (donor cells) from minipig and Fengjing pig respectively. Embryo transfer was carried out using the two kinds of reconstructed embryos. Result There were no significant differences between the two kinds of donor cells in vitro development (P>0.05). Chemical activation after electric fusion could partially improve the blastocyst rates, but the difference was not significant (P>0.05). Four healthy cloned piglets were borned after embryos transfer by the mixture of the two kinds reconstructed embryos. Microsatellite identification of cloned piglets showed whole same with their donor cells in heredity, and had nothing to do with their surrogate sows. Conclusion The method of our study group could be used to produce clone pigs.

Key words: Nuclear transfer, Clone, Minipig, Chemical activation

中图分类号:

Q95-33

戴建军, 李垚, 顾晓龙, 张廷宇, 吴彩凤, 张树山, 吴斌, 陈会兰, 陈学进, 张德福. 小型猪克隆胚的构建及胚胎移植研究[J]. 实验动物与比较医学, 2011, 31(5): 333-337.

DAI Jian-jun, LI Yao, GU Xiao-long, ZHANG Ting-yu, WU Cai-feng, ZHANG Shu-shan, WU Bin, CHEN Hui-lan, CHEN Xue-jin, ZHANG De-fu. Studies on Construction and Transfer of Cloned Minipig Embryos[J]. Laboratory Animal and Comparative Medicine, 2011, 31(5): 333-337.

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小型猪克隆胚的构建及胚胎移植研究

Studies on Construction and Transfer of Cloned Minipig Embryos

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