兔皮肤成纤维细胞的体外培养和NESTED-PCR支原体污染检测  

In Vitro Culture of Rabbit Skin Fibroblasts and Detection of Mycoplasma Contamination by Nested PCR

作　　者：杨素芳[1] 崔奎青[1] 陈华海[1] 韦精卫[1] 韦兰锋[2] 石德顺[1] 

机构地区：[1]广西大学动物繁殖研究所,广西南宁530005 [2]广西大学动物科学技术学院,广西南宁530005

出　　处：《安徽农业科学》2008年第13期5305-5306,5328,共3页Journal of Anhui Agricultural Sciences

基　　金：广西青年科学基金资助项目(桂科青0542022)

摘　　要：[目的]为动物细胞核移植、转基因操作等提供充足的供体材料。[方法]通过对兔皮肤成纤维细胞的体外原代、传代培养和冷冻复苏,检测支原体污染状况。[结果]结果表明:在含有15%FCS(Fetal Calf Serum)的DMEM中培养的细胞初期生长状态较10%FCS中的细胞长势好,后期则相反。用PCR法对培养至25代的兔皮肤成纤维细胞进行支原体检测,均没有发现支原体污染现象。[结论]PCR法检测细胞培养支原体污染具有灵敏、准确、简便、快速、特异性好的优点。[ Objective ] The research aimed to provide donor material for animal cell nuclear transfer and wansgenic operation. [ Method ] The situation of mycoplasma contamination was detected after the vitro primary culture , subculture and freezing-thawing of rabbit skin fibroblasts. [ Results ] The early growth of rabbit skin fibroblasts growed better in DMEM medium with 15% fetal calf serum than in that of 10% fetal calf serum. The situation was reverse in lately growth. There was no mycoplasma contamination found in the 25 generations of rabbit skin fibroblasts by PCR. [ Conclusion] The method of PCR sensitive, accurate, simple, fast and selective.

关 键 词：兔成纤维细胞 细胞培养 血清 支原体 

分 类 号：S821.3[农业科学—畜牧学]