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体外培养山羊成纤维细胞系方法的建立
引用本文:潘求真,田亮,徐曙光,韩红兵,李佳,李海,孙书锋,连正兴,杨宁,李宁,谭景和.体外培养山羊成纤维细胞系方法的建立[J].中国农业大学学报,2006,11(1):29-34.
作者姓名:潘求真  田亮  徐曙光  韩红兵  李佳  李海  孙书锋  连正兴  杨宁  李宁  谭景和
作者单位:1. 中国农业大学,动物科学技术学院,北京,100094;东北农业大学,生命科学学院,哈尔滨,150030
2. 中国农业大学,动物科学技术学院,北京,100094
3. 东北农业大学,生命科学学院,哈尔滨,150030
基金项目:国家自然科学基金资助项目(30170675)
摘    要:用山羊组织块和0.25%的胰蛋白酶消化培养法获得正常传代细胞,对于出现的上皮样细胞和成纤维样细胞混合生长的问题,则根据两者贴壁紧实程度不同,用0.05%的胰酶-EDTA进行不同时间的消化,将其分离纯化。纯化的成纤维体细胞经数次传代培养后,进行冷冻解冻检验表明仍具有正常的传代能力。各代体细胞的核型分析表明,在体外培养至20~30代成纤维细胞的细胞形态(为梭形细胞,高度汇合后呈火焰状)、细胞周期以及核型均为正常,符合体细胞克隆转基因的基本要求。

关 键 词:山羊  成纤维细胞  体外培养
文章编号:1007-4333(2006)01-0029-06
收稿时间:2005-12-20
修稿时间:2005年12月20

A method established for goat fibroblast in vitro
Pan Qiuzhen,Tian Liang,Xu Shuguang,Han Hongbing,Li Ji,Li Hai,Sun Shufeng,Lian Zhengxing,Yang Ning,Li Ning,Tan Jinghe.A method established for goat fibroblast in vitro[J].Journal of China Agricultural University,2006,11(1):29-34.
Authors:Pan Qiuzhen  Tian Liang  Xu Shuguang  Han Hongbing  Li Ji  Li Hai  Sun Shufeng  Lian Zhengxing  Yang Ning  Li Ning  Tan Jinghe
Abstract:Goat fibroblasts were cultured by tissue pieces,trypsin,respectively.Establish an ideal cellular culture methods.Especially,in the tissue pieces,first of all,we cut the tissue into 1?mm~3 pieces,second dry the pieces by the centrifugation,third,add a little fetal bovine serum onto pieces,and then,paved the piece onto the culture plate in even,cultured for 60-90?min.in incubator,last,add the culture medium.The two method had a 100% of primary passagey fibroblast cells present rate and successful passage rate.Two methods had a differences between primary fibroblast cells present rate and time of can be passaged.Using different digested time to purified fibroblast cells with epithelial cells.Purified fibroblast cells still have normal capacity of passage after freezethawed.Kary otype analyze and cell cycle indicate that after 20-30 passage of in vitro culture no abnormal.It suit for the need of clone and transgenic chone.
Keywords:goat  fibroblast cell  in vitro culture
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