CELL LINES AUTHENTICATION
Cell cultures are widely used in many laboratories. However, most cell lines are not correctly identified or are contaminated with other cells. This leads to doubtfulness of the authentication of the published data. It is estimated by ATCC that up to 30% of cell lines are incorrectly characterized, especially cancer cell lines, in which genetic aberrations occur often. Because of the magnitude of the problem, cell line identification is a prerequisite, for example for obtaining a grant from National Institutes of Health (NIH) and also in publications, in many scientific journals.
The Cell Line Identification is performed via DNA profiling of short tandem repeats (STR-loci) using at least eight STR-loci as recommended for standardization of the analysis: TH01, TPOX, vWA, CSF1PO, D16S539, D7S820, D13S317, D5S818 and the amelogenin locus for human cell lines sex identification.
KRISHGEN offers the DharmaPlex20 kit for STR-typing of human DNA. Nineteen STR-markers are amplified, including the eight recommended markers and human amelogenin locus.
Table 1. List of the Analysed Loci of DharmaPlex20 kit, chromosomal localization and the eight STR-markers recommended by the American Type Culture Collection (ATCC).
Our kit is validated for working with genetic analyzers ABI (series 310/3130/3500). The obtained data is analysed using GeneMapper, OSIRIS, GeneMarker software.
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)
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.