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278种广泛使用的肿瘤细胞系的交叉污染和错误鉴定调查

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines.

作者信息

Huang Yaqing, Liu Yuehong, Zheng Congyi, Shen Chao

机构信息

State Key Laboratory of Virology, College of Life Science, Wuhan University, Wuhan, Hubei, China.

China Center for Type Culture Collection, Wuhan, Hubei, China.

出版信息

PLoS One. 2017 Jan 20;12(1):e0170384. doi: 10.1371/journal.pone.0170384. eCollection 2017.

Abstract

In recent years, biological research involving human cell lines has been rapidly developing in China. However, some of the cell lines are not authenticated before use. Therefore, misidentified and/or cross-contaminated cell lines are unfortunately commonplace. In this study, we present a comprehensive investigation of cross-contamination and misidentification for a panel of 278 cell lines from 28 institutes in China by using short tandem repeat profiling method. By comparing the DNA profiles with the cell bank databases of ATCC and DSMZ, a total of 46.0% (128/278) cases with cross-contamination/misidentification were uncovered coming from 22 institutes. Notably, 73.2% (52 out of 71) of the cell lines established by the Chinese researchers were misidentified and accounted for 40.6% of total misidentification (52/128). Further, 67.3% (35/52) of the misidentified cell lines established in laboratories of China were HeLa cells or a possible hybrid of HeLa with another kind of cell line. Furthermore, the bile duct cancer cell line HCCC-9810 and degenerative lung cancer Calu-6 exhibited 88.9% match in the ATCC database (9-loci), indicating that they were from the same origin. However, when we used 21-loci to compare these two cell lines with the same algorithm, the percent match was only 48.2%, indicating that these two cell lines were different. The SNP profiles of HCCC-9810 and Calu-6 also revealed that they were different cell lines. 150 cell lines with unique profiles demonstrated a wide range of in vitro phenotypes. This panel of 150 genomically validated cancer cell lines represents a valuable resource for the cancer research community and will advance our understanding of the disease by providing a standard reference for cell lines that can be used for biological as well as preclinical studies.

摘要

近年来,涉及人类细胞系的生物学研究在中国迅速发展。然而,一些细胞系在使用前未经过鉴定。因此,不幸的是,错误鉴定和/或交叉污染的细胞系很常见。在本研究中,我们使用短串联重复序列分析方法,对来自中国28个研究所的278个细胞系进行了交叉污染和错误鉴定的全面调查。通过将DNA图谱与美国模式培养物集存库(ATCC)和德国微生物和细胞培养物保藏中心(DSMZ)的细胞库数据库进行比较,发现来自22个研究所的共46.0%(128/278)的病例存在交叉污染/错误鉴定。值得注意的是,中国研究人员建立的细胞系中有73.2%(71个中的52个)被错误鉴定,占总错误鉴定的40.6%(52/128)。此外,在中国实验室建立的错误鉴定细胞系中,67.3%(35/52)是HeLa细胞或HeLa与另一种细胞系的可能杂交体。此外,胆管癌细胞系HCCC - 9810和退行性肺癌细胞系Calu - 6在美国模式培养物集存库数据库(9个位点)中的匹配度为88.9%,表明它们来自同一来源。然而,当我们使用相同算法用21个位点比较这两个细胞系时,匹配度仅为48.2%,表明这两个细胞系不同。HCCC - 9810和Calu - 6的单核苷酸多态性(SNP)图谱也显示它们是不同的细胞系。150个具有独特图谱的细胞系表现出广泛的体外表型。这一组150个经过基因组验证的癌细胞系代表了癌症研究界的宝贵资源,将通过为可用于生物学以及临床前研究的细胞系提供标准参考,推进我们对该疾病的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/faf8/5249119/944ca23bb140/pone.0170384.g001.jpg

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