2 found 631 publications published between the years 2000–2014 that mentioned the use of the KB cell line, of which 574 articles were describing it incorrectly. For example, the KB cell line, believed to be oral or squamous cell carcinoma 2, and the KU7 cell line believed to be derived from bladder cancer cells 3, were both found to be HeLa cells. There have been reports where the cell line was misidentified by the source institute, rendering the results of any publication using that cell line questionable. Misidentified or contaminated cell lines can present irreproducible or inaccurate results which may mislead future research 1. Metagenomics-based HTS is a useful laboratory QA tool for cell line authentication and contamination detection that should be conducted regularly.Ĭell line authentication is an essential part of ensuring the validity of research and diagnostic results. However, several expected and unexpected viral sequences were detected, including part of the classical swine fever virus genome in the IB-RS-2 Clone D10 cell line. Mycoplasma contamination was not detected in any cell lines. Two cell lines, LFBK-αvβ6 and SCP-HS, were determined to be composed of cells from a different species and genus. Through sequence comparison of the cytochrome c oxidase subunit 1 (COX1) gene, the species identity of 53 cell lines was confirmed, and eight cell lines were found to show a greater pairwise nucleotide identity in the COX1 sequence of a different species within the same expected genus. Cell line authentication and contamination detection based on metagenomic high-throughput sequencing (HTS) was tested on DNA and RNA from 63 cell lines available at the Canadian Food Inspection Agency’s National Centre for Foreign Animal Disease.
Lack of cell line authentication can result in the use of contaminated or misidentified cell lines, potentially affecting the results from research and diagnostic activities. Cell lines are widely used in research and for diagnostic tests and are often shared between laboratories.
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