Northwestern University Feinberg School of Medicine

Center for Genetic Medicine

Human Cell Line Authentication through STR Profiling

Projects based on the use of cell line(s) may have a need to validate the cell line(s) being used, as their identity could have changed over time due to natural genomic alterations or contamination. For projects that start anew, the importance of authenticating the cell line(s) to be used, potentially for years to come, is apparent. The risks of using unauthenticated cell lines include adding misinformation to scientific literature, waste of funding and resources, and irreproducibility of results.

Cell line authentication (CLA) is an essential aspect of addressing the increasing concern for scientific rigor and reproducibility. This has led to the establishment of requirements for proof of cell line identity and purity by funding agencies (including NIH) and journals (including Nature journals, whole list). The #authenticate campaign launched by Global Biological Standards Institute (GBSI) put the focus on the importance of CLA and has led to the establishment of Cell Authentication Alliance with cosignatories including a number of nonprofit funding agencies.   

There is an array of methods for testing cell line vis-à-vis species, chromosomal structural changes, spontaneous mutations/genetic drift, contamination with adventitious agents, and interspecies/intraspecies contamination. These methods include DNA-based species barcoding, karyotyping, profiling of STRs (Short Tandem Repeats), genotyping of SNPs, PCR amplification using species-specific primers, and whole-genome sequencing. While WGS will eventually provide thorough evaluation of a cell line, STR profiling is by far the most developed and cost-effective approach for authenticating human cell lines (details on STR profiling). Similar to forensic DNA identification, assaying a relatively small number of STR loci provides enough power to discriminate different human cell lines.

NUSeq uses a commercially available kit (AmpFLSTR® Identifiler® Plus PCR Amplification Kit) for STR profiling. This is the same kit used by forensic scientists for casework. This kit is a multiplex STR assay allowing amplification of 15 tetranucleotide repeat loci and the Amelogenin marker (for gender determination) in a single PCR amplification.

Human Cell Line Authentication Pricing

Human cell line authentication costs are listed on the Core Pricing page.

Human Cell Line Authentication