CELL LINES

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.

Locus Chromosome ATCC recommended DharmaPlex20
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 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)
http://www.dsmz.de/services/services-human-and-animal-cell-lines/online-str-analysis.html

Cell Line Integrated Molecular Authentication (CLIMA)
http://bioinformatics.istge.it/clima/

American Type Culture Collection (ATCC)
https://www.atcc.org/CulturesandProducts/CellBiology/STRProfileDatabase/tabid/174/Default.aspx

Japanese Collection of Research Bioresource (JCRB)
http://cellbank.nibio.go.jp/english/

Literature

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.

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