Human Cell Line and Tissue Sample Authentication

Background: Short Tandem Repeat (STR) genotyping analysis is a proven technology for uniquely identifying virtually all human samples. STR genotyping was adopted as the preferred technology for identification of human tissue culture cell lines by the ATCC Standards Development Organization (ASN-0002: Authentication of Human Cell Lines: Standardization of STR Profiling). We developed new automation-compatible protocols/systems for generating STR profiles from human cell lines or tissue samples. Methods: We adapted the STR genotyping systems routinely used for forensic and paternity testing to better meet the needs of genomic core facilities. Modifications include balancing for higher amounts of template DNA, configuring the reagents for compatibility with high throughput robotic workstations, and supporting electrophoretic separation and analysis on a wider variety of instrument and software platforms. Results: The GenePrint® 10 and GenePrint® 21 Systems allow for multiplexed genotyping of 9 or 20 STR loci, plus the amelogenin gender marker. STR analysis with these loci uniquely identify virtually all human cell lines and tissue samples (body fluids, tissues and extracted DNA) and confirm the absence of cross-contamination or a sample switch. The GenePrint® 10 System is compatible with purified DNA or direct amplification from cells deposited on FTA® Cards (GE Whatman). Both genotyping systems meet the ASN-0002 standard and include the loci represented on the National Center for Biotechnology Information (NCBI) human cell line database: http://www.ncbi.nlm.nih.gov/biosample?term=human%20cell%20line%20STR%20profile Conclusions: STR genotyping analysis with the GenePrint® 10 and GenePrint® 21 Systems can establish human cell line or tissue sample identity and confirm the absence of contamination with other human cell lines or tissues. The methods are compatible with DNA concentrations, robotic protocols, instrumentation, and genotyping software typically used in genomics core facilities.