应用聚合酶链式反应鉴定实验细胞的来源种属  

Species identification of animal cells by polymerase chain reaction

作　　者：卞晓翠[1] 刘玉琴[1] 王春景[1] 苏小玲[1] 赵晓梅[1] 顾蓓[1] 张宏[1] 

机构地区：[1]中国医学科学院北京协和医学院基础医学研究所细胞中心,北京100005

出　　处：《基础医学与临床》2009年第4期423-430,共8页Basic and Clinical Medicine

基　　金：国家科技基础条件平台建设基金(20051KA21502)

摘　　要：目的应用聚合酶链式反应(PCR)鉴定常见细胞的种属来源,以判断培养细胞是否存在种间的交叉污染。方法根据文献报道和NCBI数据库我们得到了32对种属特异性引物,并分别针对10种常见的细胞种属,对这些引物进行特异性和敏感性的筛选;以待检测细胞的基因组DNA为模板进行PCR及后续的琼脂糖凝胶电泳;以不同种属来源细胞的DNA混合物作为阳性对照模板,以水作为阴性对照模板。结果针对上述10种常见细胞来源的种属分别得到了1对特异性和敏感性均较好的引物,经PCR扩增和琼脂糖凝胶电泳后可以准确鉴定待检测细胞的种属来源,并判断该细胞是否存在种间污染。结论应用聚合酶链式反应可以简便、快速地鉴定实验细胞的种属,并判断该细胞是否存在种间的交叉污染。Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction （PCR）. Methods From references and NCBI database, we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells. Then we screened for optional primers with high specificity and high sensitivity. PCR amplification with these primers, the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR pr-control template, and water as negative-control templateoducts were done. Mixed DNA of 10 species was used as positive. Results Ten pairs of species-specific and highly sensitive primers were selected. By PCR amplification with these primers and agarose gel electrophoresis, we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species. Conclusion This PCR assay provides a simple, rapid, sensitive, and cost-effective method to identify cell species and detect interspecies cross-contamination.

关 键 词：聚合酶链式反应 细胞培养 种属鉴定 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]