基础医学与临床 ›› 2009, Vol. 29 ›› Issue (4): 423-430.

• 技术与方法 • 上一篇    下一篇

应用聚合酶链式反应鉴定实验细胞的来源种属

卞晓翠 刘玉琴 王春景 苏小玲 赵晓梅 顾蓓 张宏   

  1. 中国医学科学院基础医学研究所;北京协和医学院基础学院 中国医学科学院基础医学研究所;北京协和医学院基础学院 中国医学科学院基础医学研究所;北京协和医学院基础学院 中国医学科学院基础医学研究所;北京协和医学院基础学院 中国医学科学院基础医学研究所;北京协和医学院基础学院
  • 收稿日期:2008-09-05 修回日期:2008-10-09 出版日期:2009-04-25 发布日期:2009-04-25
  • 通讯作者: 刘玉琴

Species identification of animal cells by polymerase chain reaction

Xiao-cui BIAN, Yu-qin LIU, Chun-jing WANG, Xiao-ling SU, Xiao-mei ZHAO, Bei GU, Hong ZHANG   

  1. Cell Culture Center, IBMS, CAMS & PUMC Cell Culture Center, IBMS, CAMS & PUMC Cell Culture Center, IBMS, CAMS & PUMC Cell Culture Center, IBMS, CAMS & PUMC Cell Culture Center, IBMS, CAMS & PUMC
  • Received:2008-09-05 Revised:2008-10-09 Online:2009-04-25 Published:2009-04-25
  • Contact: Yu-qin LIU,

摘要: 目的 应用聚合酶链式反应(PCR法)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。

关键词: 聚合酶链式反应, 细胞培养, 种属鉴定

Abstract: 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 commercially synthesized 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells. Then we screened for optional primers with good specificity and high sensitivity. PCR amplification with these primers and the genomic DNAs isolated from the tested cell line, and agarose gel electrophoresis of the PCR products were done. Mixed DNA of 10 species was used as positive-control template, and water as negative-control template. Results Finally 10 pairs of species-specific and high sensitive primers were selected. By PCR amplification with these primers and agarose gel electrophoresis, we could easily tell the origin of cell lines and detect whether the tested cell lines are contaminated by cells of other species. Conclusions This PCR assay provides a simple, rapid, sensitive, and cost-effective method to identify cell species and detect interspecies cross-contamination.

Key words: polymerase chain reaction, cell culture, species identification