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CELL LINE MISIDENTIFICATION

“Evidence suggests that up to one-third of tumor cell lines being used in scientific research are affected by inter- or intraspecies cross-contamination or have been wrongly identified, thereby rendering many of the conclusions doubtful if not completely invalid.” —  Lancet Oncology, vol. 2, July 2001, p. 393

Tools to Prevent Cell Line Contamination and Misidentification

Simple pipetting and labeling errors can happen to the most conscientious cell culturist and lead to the propagation of a misidentified cell line. Additionally, both intra- and interspecies cell line cross-contamination can lead to misidentification if the contaminating cell line is heartier than the cell line it has corrupted. Proper aseptic technique can reduce the risk of contamination and misidentification, but nobody is perfect - and the potential cost of using contaminated or misidentified lines, in time, resources, and reputation, is very high. Thus, periodic authentication of your cell lines is well-worth the effort.

Cell lines from ATCC have been thoroughly tested and authenticated, so you can be certain the label on the vial represents the cells stored within. If, however, your cells come from a less than reliable source or if they’ve been sharing an incubator with other cell lines for a while, it may be time to perform an identity check.

ATCC uses morphology, karyotyping, and PCR based approaches to confirm the identity of human cell lines and to rule out both intra- and interspecies contamination. PCR based methods include, an assay to detect species specific variants of the cytochrome C oxidase I gene (COI analysis) to rule out inter-species contamination and short tandem repeat (STR) profiling to distinguish between individual human cell lines and rule out intra-species contamination.

Check out the resources listed below for more information, and for a description of the tools ATCC uses to ensure the cell lines in the collection are properly identified and free from contamination.

Misidentification of human cell lines: Science vs. Policy – 2012
Dr. Yvonne Reid, a cell authentication specialist at ATCC, gave this presentation at the European Stem cell meeting held in San Diego, CA, this past February. Her talk describes the history of the problem and outlines how scientific policy can help to eradicate cell contamination and misidentification.

Development of a Consensus Standard for the Authentication of Human Cell lines: Standardization of STR Profiling – Part I: History
Dr. John Masters, of the University College London, provides an historical overview of the problem, highlights some examples of cell lines that are known to be contaminated or misidentified, and illustrates how these “false” lines have permeated the scientific literature.

Check your cultures! A list of cross-contaminated or misidentified cell Lines (Capes-Davis et al, 2010)
The authors of this study reviewed the publically available information on PubMed, Wikipedia, the website of the ATCC, and the websites of other cell repositories to develop a comprehensive list of cell lines known to be contaminated or misidentified.

ATCC uses morphology, karyotyping, and PCR based approaches to confirm the identity of human cell lines and to rule out both intra- and interspecies contamination. PCR based methods include, an assay to detect species specific variants of the cytochrome C oxidase I gene (COI analysis) to rule out inter-species contamination and short tandem repeat (STR) profiling to distinguish between individual human cell lines and rule out intra-species contamination.

Morphology

COI Analysis

Karyotype

STR Profiling

See how ATCC combats cell  misidentification