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Cloned goats(Capra hircus)from adult ear cells
作者姓名:郭继彤  安志兴  李煜  李雪峰  李裕强  郭泽坤  张涌
作者单位:GUO Jitong,AN Zhixing LI YuLI Xuefeng,LI Yuqiang GUO Zekun& ZHANG Yong1. College of Life Science,Inner Mongolia University,Huhhot 010021,China;2. Institute of Bioengineering,Northwest Sci-tech University of Agricultu
基金项目:This work was finished in Northwest Sci-tech University of Agriculture and Forestry. We thank Prof. Chen Sumin, Chen Nanchun and Dr. Chai Yubo for microsatellite DNA analysis. And we thank Dr. Wang Xinzhuang, Liu Zelong for help of embryos transfer in g
摘    要:The average number of available oocytes recovered per ovary collected during the breeding season in dairy goats was 5.5 (1815/330). 66.17% (1201/1815) of oocytes extruded the first polar body after maturation in vitro for 20 h. 75.44% (906/1201) of matured oocytes with membrane evagination around the MII chromosomes were enucleated. Ear skin fibroblast cells were derived from an adult female dining Grey goat (C. hircus). The cells were cryopreserved in liquid nitrogen after passage 2. Thawed cells were further cultured for 3-6 passages and were subjected to serum starvation by 0.5% FBS for 2-10 d, then used as donor cells for nuclear transfer. 98.12% (889/906) of the enucleated oocytes were reconstructed by intracytoplasmic injection of karyoplast. The reconstructed embryos were activated by 5μ mol/L ionomycin for 4.5 min and further activated by culturing with 6-dimethylaminopurine (6-DMAP) for 3 h. After 36 h of culture in mCR1aaBF, 76.69% (645/841) of the cloned embryos cleaved. There were no signifi

收稿时间:2001-07-20

Cloned goats (Capra hircus) from adult ear cells
Guo Jitong,An Zhixing,Li Yu,Li Xuefeng,Li Yuqiang,Guo Zekun,Zhang Yong.Cloned goats(Capra hircus)from adult ear cells[J].Science China Life sciences,2002,45(3):260-267.
Authors:Guo Jitong  An Zhixing  Li Yu  Li Xuefeng  Li Yuqiang  Guo Zekun  Zhang Yong
Institution:College of Life Science, Inner Mongolia University, 010021, Huhhot, China, gjitong@hotmail.com.
Abstract:The average number of available oocytes recovered per ovary collected during the breeding season in dairy goats was 5.5 (1815/330). 66.17% (1201/1815) of oocytes extruded the first polar body after maturation in vitro for 20 h. 75.44% (906/1201) of matured oocytes with membrane evagination around the M II chromosomes were enucleated. Ear skin fibroblast cells were derived from an adult female Jining Grey goat (C. hircus). The cells were cryopreserved in liquid nitrogen after passage 2. Thawed cells were further cultured for 3-6 passages and were subjected to serum starvation by 0.5% FBS for 2-10 d, then used as donor cells for nuclear transfer. 98.12% (889/906) of the enucleated oocytes were reconstructed by intracytoplasmic injection of karyoplast. The reconstructed embryos were activated by 5 mumol/L ionomycin for 4.5 min and further activated by culturing with 6-dimethylaminopurine (6-DMAP) for 3 h. After 36 h of culture in mCR1aaBF, 76.69% (645/841) of the cloned embryos cleaved. There were no significant differences in development in vitro between the cloned embryos derived from donor cells precooled at 4 degrees C for 24 h and nonprecooled donor cells. The cleavage rates, 4-cell development, and blastocyst development of reconstructed embryos were 72.48% (79/109), 53.16% (42/79), and 19.05% (8/42) in precooled group; 68.5% (211/308), 59.72% (126/211), and 17.46% (22/126) in nonprecooled group, respectively. Eighteen cloned 4-cell embryos derived from precooled donor cells were transferred and one cloned kid was born. Eighty-four cloned 4-cell embryos derived from nonprecooled donor cells were transferred and no offspring were produced. Of 18 cloned morale from nonprecooled donor cells transferred, one kid was born. The results of microsatellite DNA analyses indicated that the two cloned kids were from the same donor fibroblast cell line derived from an adult goat ear skin.
Keywords:goat  ear skin fibroblast cells  intracytoplasmic injection  nuclear transfer  cloned goats  
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