Cell Line Identification and Authentication: Human Cell Lines Standards and Protocols

As a prerequisite for manuscript submission and funding, many journals and funding agencies now recommend or require cell line authentication.   In the National Institute of Health’s (NIH) Notice Number: NOT-OD-15-103 "Enhancing Reproducibility through Rigor and Transparency,” released June 9, 2015, NIH revised its grant application instructions to include regular authentication of key biological and/or chemical resources.  Specifically, the agency stated that, beginning in January 2016, “NIH expects that key biological and/or chemical resources will be regularly authenticated to ensure their identity and validity for use in the proposed studies.”

The current gold standard for human cell line authentication is Short Tandem Repeats (STR) genotyping.  ANSI/ATCC consensus standard ASN-0002 “Authentication of Human Cell Lines: Standardization of STR Profiling” details all aspects of human cell line authentication using STR analysis.  Commercial kits are available for genotyping; various companies offer genotyping services for human cell line authentication, as well. 

National repositories list at least eight (8) human STR markers and amelogenin (AMEL), the sex determination marker for each human cell line deposited.  Human STR profile databases are available for human cell line data comparison.  There is also a database of misidentified and contaminated human cell lines which lists approximately 475 cell lines that are known to be misidentified.