Cell Line Authentication

Cell lines are fundamental research tools that serve as models for complex biological systems, especially those of human tissue. In order to maintain meaningful tissue-specific conclusions in cell-based experiments, as well as to ensure reproducibility, it is imperative that cell lines accurately represent the cellular identities of its original source or donor. Hepatic cell lines, for instance, must comprise of liver cells. There are unfortunate instances, however, when this turns out not to be so. Lines such as Chang liver cells, have emerged to be merely a propagation of the highly robust and contaminating HeLa cells. This is just one example of the many cell lines exposed since Gartler and Nelson-Rees first began to uncover HeLa cross-contaminations in the 1960’s. To date, over 450 misidentified cell lines have since been registered by the International Cell Line Authentication Committee (ICLAC).¹ In response, an increasing number journals have begun to set guidelines for authenticating cell lines in prior to publication. As of 2016, the National Institutes of Health has announced its expectations of cell line authentication when considering grant applications.²

A widely used method for cell line authentication involves STR profiling, a PCR-based technique that compares multiple short tandem repeat (STR) markers between two or more cellular genomes. In identifying human cell lines (derived from different donors), a minimum of 8 core STR markers are needed: D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX, CSF1PO, along with an additional gender-distinguishing gene, amelogenin.³ In 2011, a collaboration between the American National Standards Institute (ANSI) and American Type Culture Collection (ATCC) has led to the creation of over 100-page documentary standard ASN-0002, providing best practice standard procedures for DNA extraction, STR profiling, data analysis and interpretation, quality control, and use of searchable databases.^4,5 Beyond STR, other methods for cell line identification can include whole genome sequencing, species-specific primers, cytochrome oxidase (CO1) profiling, and SNP analysis. Mycoplasma, another common source of contamination, and can be detected by methods such as staining, culturing, or PCR.

References:

1. ICLAC. Register of Misidentified Cell Lines. ICLAC (2016). Available at: https://iclac.org/databases/cross-contaminations/. (Accessed: 21st September 2017)
2. National Institutes of Health. Rigor and Reproducibility. (2017). Available at: https://grants.nih.gov/reproducibility/index.htm. (Accessed: 21st September 2017)
3. Reid, Y., Storts, D., Riss, T. & Minor, L. in Assay Guidance Manual (eds. Sittampalam, G. S. et al.) (Eli Lilly & Company and the National Center for Advancing Translational Sciences, 2004). Available from: https://www.ncbi.nlm.nih.gov/books/NBK144066/
4. Almeida, J. L., Cole, K. D. & Plant, A. L. Standards for Cell Line Authentication and Beyond. PLoS Biol 14, (2016).
5. ANSI. ANSI/ATCC ASN-0002-2011. Available at: https://webstore.ansi.org/RecordDetail.aspx?sku=ANSI%2FATCC+ASN-0002-2011. (Accessed: 21st September 2017)