Overview

​Cultured cells used in research can be misidentified or can become altered, either by contamination from other cell lines or by spontaneous genetic changes after a high number of subcultures. The American Type Culture Collection (ATCC®) recommends using low passage cell lines and authentication of cell lines used in research to assure reliable and reproducible results. Low passage minimizes the number of spontaneous changes that arise. Authentication provides a means to:
  • verify that the cell line is human (mouse, rat, hamster, rhesus, bovine, and fish DNA will not generate a genotype with the authentication method that is employed).
  • verify whether the genotype of the cell line is identical to the genotype of the presumed parental cells. 
  • check whether your cell line is cross-contaminated with another cell line.
One method that ATCC® suggests for verifying human cell line identity is by DNA fingerprinting by small tandem repeat (STR) profiling. The STR profiles for many human cell lines used in research are available on several public websites. (Links to these websites are available on the Results tab.) These STR profiles can be used as a reference to compare the STR genotypes of  cells submitted for analysis. If no STR profile is available for a cell line of interest, it would be prudent to DNA fingerprint the cell line to establish a reference profile for future identity verifications.   

The Molecular Diagnostics Laboratory has adopted an American National Standards Institute (ANSI) recommended protocol to authenticate human cell lines as a service to investigators. The Promega GenePrint® 10 system uses a multiplex PCR approach to coamplify 9 highly polymorphic STR markers and the Amelogenin gene in the human genome. The amplified DNA fragments of each sample are resolved on the Applied Biosystems 3130xl 16-capillary Genetic Analyzer with Applied Biosystems GeneMapper v 4.0 software to determine cell line genotypes.

*Please note that this genotyping method assays 10 human loci in the genome. Changes that may have arisen at other loci during cell culture will not be detected by this method.