| 山羊类ES细胞的分离与培养 |
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| 引用本文： | 刘艳,高其双,黄海军,邓兵,蒋思文.山羊类ES细胞的分离与培养[J].畜牧兽医学报,2011,42(8). |
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| 作者姓名： | 刘艳 高其双 黄海军 邓兵 蒋思文 |
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| 作者单位： | 1. 华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉,430070
2. 武汉市畜牧兽医科学研究所,武汉,430065 |
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| 基金项目： | 国家“863”计划(2007AA100505) |
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| 摘 要： | 旨在优化并建立有利于山羊类ES细胞生长的饲养层细胞、培养基、细胞因子和传代方法。本研究选择武汉市本地白山羊配种6～7 d后的胚胎,通过全胚培养法、酶消化与机械分离结合法分离山羊类ES细胞,并在不同的细胞饲养层中饲养、在培养基中添加不同的生长因子、采用不同的传代方法,比较不同培养条件对山羊类ES细胞生长和增殖的影响。通过碱性磷酸酶染色和免疫组化染色法鉴定ES细胞特异生物标记AKT、SSEA-1和Oct-4的表达,并将得到的山羊类ES细胞进行体外分化。结果表明,与小鼠胎儿成纤维细胞(Mouse embryonic fibro-blast,MEF)和山羊胎儿成纤维细胞(Goat embryonic fibroblast,GEF)相比,C2C12更有利于细胞的贴壁,但差异不显著(P>0.05);细胞传代时,在高浓度细胞因子LIF(Leukemia inhibitory factor)和SCF(Stemcell factor)中处理ICM(Inner cell mass),更有利于细胞传代(传至第8代);培养基中添加LIF和SCF的同时,添加肝素和胰岛素,更有利于细胞的传代(传至第9代);山羊类ES细胞中SSEA-1(...
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| 关 键 词： | 胚胎干细胞 内细胞团 山羊 |
Isolation and Culture of Goat ES-like Cells |
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| LIU Yan,GAO Qi-shuang,HUANG Hai-jun,DENG Bing,JIANG Si-wen.Isolation and Culture of Goat ES-like Cells[J].Acta Veterinaria et Zootechnica Sinica,2011,42(8). |
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| Authors: | LIU Yan GAO Qi-shuang HUANG Hai-jun DENG Bing JIANG Si-wen |
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| Institution: | LIU Yan1,GAO Qi-shuang2,HUANG Hai-jun2,DENG Bing1,JIANG Si-wen1 (1.Key Laboratory of Agricultural Animal Genetics,Breeding and Reproduction of Ministry of Education,Huazhong Agricultural University,Wuhan 430070,China,2.Wuhan Institute of Animal and Veterinary Science,Wuhan 430065,China) |
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| Abstract: | The aim of this study was to optimize and set up better goat feeder layer cell,culture medium,cell factors and passaging method for better growth of goat ES-like cells.Blastocysts obtained by whole embryo cultured,trypsin digestion and mechanical dissociation after enzymatic treatment from Wuhan goats 6-7 days after mating were cultivated.The effect of different cell layers,different cell factors in culture medium and different passaging methods on the cell survival and differentiation were compared.Alkalin... |
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| Keywords: | embryonic stem cell inner mass cell goat |
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