Korch Christopher, Hall Erin M, Dirks Wilhelm G, Ewing Margaret, Faries Mark, Varella-Garcia Marileila, Robinson Steven, Storts Douglas, Turner Jacqueline A, Wang Ying, Burnett Edward C, Healy Lyn, Kniss Douglas, Neve Richard M, Nims Raymond W, Reid Yvonne A, Robinson William A, Capes-Davis Amanda
International Cell Line Authentication Committee (ICLAC).
Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora CO.
Int J Cancer. 2018 Feb 1;142(3):561-572. doi: 10.1002/ijc.31067. Epub 2017 Oct 10.
A variety of analytical approaches have indicated that melanoma cell line UCLA-SO-M14 (M14) and breast carcinoma cell line MDA-MB-435 originate from a common donor. This indicates that at some point in the past, one of these cell lines became misidentified, meaning that it ceased to correspond to the reported donor and instead became falsely identified (through cross-contamination or other means) as a cell line from a different donor. Initial studies concluded that MDA-MB-435 was the misidentified cell line and M14 was the authentic cell line, although contradictory evidence has been published, resulting in further confusion. To address this question, we obtained early samples of the melanoma cell line (M14), a lymphoblastoid cell line from the same donor (ML14), and donor serum preserved at the originator's institution. M14 samples were cryopreserved in December 1975, before MDA-MB-435 cells were established in culture. Through a series of molecular characterizations, including short tandem repeat (STR) profiling and cytogenetic analysis, we demonstrated that later samples of M14 and MDA-MB-435 correspond to samples of M14 frozen in 1975, to the lymphoblastoid cell line ML14, and to the melanoma donor's STR profile, sex and blood type. This work demonstrates conclusively that M14 is the authentic cell line and MDA-MB-435 is misidentified. With clear provenance information and authentication testing of early samples, it is possible to resolve debates regarding the origins of problematic cell lines that are widely used in cancer research.
多种分析方法表明，黑色素瘤细胞系UCLA - SO - M14（M14）和乳腺癌细胞系MDA - MB - 435源自同一个供体。这表明在过去的某个时间点，其中一个细胞系被错误识别，即它不再与报告的供体相对应，而是被错误地认定（通过交叉污染或其他方式）为来自不同供体的细胞系。初步研究得出结论，MDA - MB - 435是被错误识别的细胞系，M14是真实的细胞系，尽管有相互矛盾的证据发表，这导致了进一步的混乱。为了解决这个问题，我们获取了黑色素瘤细胞系（M14）的早期样本、来自同一供体的淋巴母细胞系（ML14）以及保存在原始机构的供体血清。M14样本于1975年12月冷冻保存，早于MDA - MB - 435细胞系在培养中建立的时间。通过一系列分子特征分析，包括短串联重复序列（STR）分型和细胞遗传学分析，我们证明后来的M14和MDA - MB - 435样本与1975年冷冻的M14样本、淋巴母细胞系ML14以及黑色素瘤供体的STR图谱、性别和血型相对应。这项工作确凿地证明M14是真实的细胞系，而MDA - MB - 435被错误识别。有了明确的来源信息和早期样本的鉴定测试，就有可能解决关于癌症研究中广泛使用的有问题细胞系起源的争议。
