该研究旨在鉴定国内实验室肺癌细胞系使用的正确性,对从国内多个实验室收集的54例人肺癌细胞样品提取基因组DNA,使用PCR技术扩增短串联重复序列(short tandem repeat,STR)后进行毛细管电泳,获得细胞的STR图谱.与国际数据库中已知细胞系的STR图谱进行比对,通过计算匹配度确认细胞系的身份信息,判断细胞是否存在交叉污染.结果表明54例肺癌细胞样品中存在交叉污染的有14例,错误率为25.93％(14/54).其中51例常见人肺癌细胞样品的错误率为27.45％(14/51),3例中国科学院分子细胞科学卓越创新中心/生物化学与细胞生物学研究所科学家建立的人肺癌细胞系样品的错误率为0％(0/3).该研究针对国内细胞系交叉污染的情况,分析了其原因并提出了相应的建议.
Analysis of Cross-Contamination in 54 Human Lung Cancer Cell Lines
This study aimed to authenticate the correctness of human lung cancer cell lines commonly used in Chinese labs.Genomic DNA was extracted from 54 human lung cancer cell samples collected from several domestic labs,STR(short tandem repeat)sequences were amplified by PCR,and capillary electrophoresis was performed to obtain the STR profiles of these cells.When compared to their known STR profiles released in the international databases,the identity information of these cell lines was confirmed by the matching rate,and cell cross-contamination was determined.The results showed that 14 of the 54 cell lines were cross-contaminated,with an error rate of 25.93％(14/54).The error rate of 51 common human lung cancer cell samples was 27.45％(14/51),and the error rate of three samples of human lung cancer cell lines established by the scientists in Shanghai Institute of Biochemistry and Cell Biology,Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences was 0％(0/3).The causes of cross-contamination of cell lines in Chinese labs were analyzed,and solutions for this serious phenomenon were proposed.
short tandem repeat typinghuman lung cancer cell linecross-contamination
NETL
NSTL
万方数据
