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化学辅助去核法对绵羊卵母细胞去核率和重构胚发育率的影响
引用本文:潘晓燕,王正朝,李质馨,金玉姬,窦肇华,郭志勤,王锋.化学辅助去核法对绵羊卵母细胞去核率和重构胚发育率的影响[J].生物工程学报,2009,25(4):503-508.
作者姓名:潘晓燕  王正朝  李质馨  金玉姬  窦肇华  郭志勤  王锋
作者单位:1. 吉林医药学院,组织学与胚胎学教研室,吉林,132013
2. 弗吉尼亚联邦大学医学院,里士满,23298
3. 新疆畜牧科学院,农业部家畜繁育生物技术重点开放实验室,乌鲁木齐,830000
4. 南京农业大学,动物胚胎工程技术中心,南京,210095
基金项目:吉林医药学院科研专项基金(No.2008-01);;新疆维吾尔自治区重点科技攻关项目(No.200631109)资助~~
摘    要:为提高绵羊体细胞核移植的效率,本研究采用一种新的去核方法—化学辅助去核法,对绵羊体外成熟的卵母细胞进行去核,研究了化学诱导剂秋水仙素的处理浓度、作用时间、卵母细胞的成熟时间对去核效果及重构胚发育的影响。结果表明:1)卵母细胞在0.4μg/mL的秋水仙素溶液中分别孵育0.5h和1h,胞质突起率和去核率没有显著的差异,突起率可高达85.4%,去核率达到100%;2)0.2μg/mL或0.4μg/mL秋水仙素溶液将卵母细胞处理0.5h,对去核效果没有显著影响;3)对于体外成熟18~23h的卵母细胞,随着成熟时间的延长,盲吸法的去核率降低,但没有影响秋水仙素诱导胞质突起的比率和去核率;4)两种去核方法对重构胚的发育没有产生显著影响,但成熟21~23h卵母细胞重构胚囊胚的发育率显著高于成熟18~20h卵母细胞重构胚囊胚的发育率。综上所述,本试验优化了绵羊卵母细胞化学辅助的去核程序,利用化学辅助去核法对高卵龄的绵羊卵母细胞进行去核,提高了去核率和重构胚的体外发育率。

关 键 词:秋水仙素  卵母细胞  化学辅助去核  重构胚  
收稿时间:9/7/2008 12:00:00 AM

Effect of the chemically assisted enucleation on the enucleation of sheep oocytes and the development of their reconstructed embryos
Xiaoyan Pan,Zhengchao Wang,Zhixin Li,Yuji Jin,Zhaohua Dou,Zhiqin Guo and Feng Wang.Effect of the chemically assisted enucleation on the enucleation of sheep oocytes and the development of their reconstructed embryos[J].Chinese Journal of Biotechnology,2009,25(4):503-508.
Authors:Xiaoyan Pan  Zhengchao Wang  Zhixin Li  Yuji Jin  Zhaohua Dou  Zhiqin Guo and Feng Wang
Institution:Department of Histology and Embryology, Jilin Medical College, Jilin 132013, China;Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298, USA;Department of Histology and Embryology, Jilin Medical College, Jilin 132013, China;Department of Histology and Embryology, Jilin Medical College, Jilin 132013, China;Department of Histology and Embryology, Jilin Medical College, Jilin 132013, China;Key Laboratory of Animal & Veterinary Biotechnology of MOA, Urumqi 830000, China;Center of Animal Embryo Engineering and Technology, Nanjing Agricultural University, Nanjing 210095, China
Abstract:In order to enhance the efficiency of sheep somatic cell nuclear transfer,we used a chemically assisted enucleation with colchicine to study the effects of the concentration of colchicine,the incubation time of oocytes in colchicine and the maturation time of oocytes on the enucleation rates and the development of reconstructed embryos. The results showed that 1) there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 ...
Keywords:colchicine  oocyte  chemically assisted enucleation  reconstructed embryos  
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