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Cell line authentication

Cell Lines Authentication

Cell culture is widely used in many laboratories. Unfortunately, most cell lines are not correctly identified or contaminated with other cells. This often leads to disproof of the already published data, wasted time and reagents. This problem is very widespread – up to 25% of cell lines are incorrectly characterized, especially cancer cell lines, in which genetic aberrations occur often [1-3]. Because of the magnitude of the problem, cell line identification is required, for example for obtaining a grant from National Institutes of Health (NIH) and publication in many scientific journals. The identification is performed via DNA profiling of short tandem repeats (STR-loci), moreover, using of at least eight STR-loci is recommended for standardization of the analysis [4]: TH01, TPOX, vWA, CSF1PO, D16S539, D7S820, D13S317, D5S818 and the amelogenin locus for human cell lines’ sex identification. Gordiz company has developed and manufactures COrDIS Plus kit for STR-typing of human DNA. Nineteen STR-markers are amplified, including the eight recommended markers and human amelogenin locus.

Locus Chromosome ATCC recommended
Amelogenin X Xp22.1-22.3 + +
Amelogenin Y Yp11.2
D3S1358 3p21.31   +
TH01 11p15.5 + +
D12S391 12p13.2   +
D1S1656 1q42   +
D10S1248 10q26.3   +
D22S1045 22q12.3   +
D2S441 2p14   +
D7S820 7q21.11 + +
D13S317 13q31.1 + +
FGA 4q31.3   +
TPOX 2p25.3 + +
D18S51 18q21.33   +
D16S539 16q24.1 + +
D8S1179 8q24.13   +
CSF1PO 5q33.1 + +
D5S818 5q23.2 + +
VWA 12p13.31 + +
D21S11 21q21.1   +
SE33 6q14   +

Table 1. List of the analyzed loci of COrDIS Plus kit, chromosomal localization and the eight STR-markers recommended by the American Type Culture Collection ATCC.


Using COrDIS Plus kit you can analyze cell lines for nineteen STR-markers and human amelogenin locus with a probability of the accidental coincidence of 10-22. COrDIS Plus kit is validated for working with genetic analyzers ABI (series 310/3130/3500) or NANOFOR 05. The obtained data is analyzed using GeneMapper, OSIRIS, GeneMarker software. Cell lines identification must be carried out periodically.

List of data bases:

German Collection of Microorganisms and Cell Cultures (DSMZ)
Cell Line Integrated Molecular Authentication (CLIMA)
American Type Culture Collection (ATCC)
Japanese Collection of Research Bioresource (JCRB)
База данных STR профилей клеточных линий NCBI BioSample

1.    Hughes, P., Marshall, D., Reid, Y., et al.  The costs of using unauthenticated, over-passaged cell lines:  how much more data do we need? Biotechniques 2007, 43, 575-586.
2.    Capes-Davis A, Theodosopoulos G, Atkin I, Drexler HG, Kohara A, Macleod RA, Masters JR, Nakamura Y, Reid YA, Reddel RR, Freshney RI (2010) Check your cultures! A list of cross-contaminated or misidentified cell lines.  Int J Cancer 127: 1-8.
3.    Masters,J.R. 2012. Cell-line authentication: End the scandal of false cell lines. Nature 492:186
4.    ANSI/ATCC ASN-0002-2011. Authentication of human cell lines: Standardization of STR profiling. ANSI eStandards Store 2012. http://webstore.ansi.org/RecordDetail.aspx?sku=ANSI%2FATCC+ASN-0002-2011