Cross-contamination of a UROtsa stock with T24 cells--molecular comparison of different cell lines and stocks

Autor: Jörg Hippler, P. Rozynek, Daniel G. Weber, Yvonne von der Gathen, O′Brien Igwilo-Okuefuna, Thomas Brüning, Elke Dopp, Irina Raiko, Ricarda Zdrenka, Georg Johnen, Oleksandr Bryk, Christian Johannes
Rok vydání: 2013
Předmět:
lcsh:Medicine
medicine.disease_cause
Molecular level
RNA interference
Molecular cell biology
Basic Cancer Research
lcsh:Science
Antigens
Viral
Tumor
Multidisciplinary
Cell immortalization
Bladder Cancer and Urothelial Neoplasias of the Urinary Tract
Cell Differentiation
Physik (inkl. Astronomie)
Bladder Cancer
Oncology
DNA methylation
Medicine
Epigenetics
DNA modification
Biologie
Research Article
Urothelial Cell
Urology
Chemie
Biology
Real-Time Polymerase Chain Reaction
Cell Line
Cell Line
Tumor
medicine
Genetics
Cancer Genetics
Humans
RNA
Messenger
lcsh:R
Cancers and Neoplasms
Sequence Analysis
DNA
DNA Contamination
DNA Methylation
Molecular biology
MicroRNAs
Genitourinary Tract Tumors
Cell culture
Cancer research
lcsh:Q
Gene expression
Urothelium
Carcinogenesis
Microsatellite Repeats
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 8, Iss 5, p e64139 (2013)
ISSN: 1932-6203
Popis: Background:UROtsa is an authentic, immortalized human urothelial cell line that is used to study the effects of metals and other toxic substances, mostly in the context of bladder cancer carcinogenesis. Unusual properties on the molecular level of a provided UROtsa cell line stock prompted us to verify its identity.Methods:UROtsa cell line stocks from different sources were tested on several molecular levels and compared with other cell lines. MicroRNA and mRNA expression was determined by Real-Time PCR. Chromosome numbers were checked and PCR of different regions of the large T-antigen was performed. DNA methylation of RARB, PGR, RASSF1, CDH1, FHIT, ESR1, C1QTNF6, PTGS2, SOCS3, MGMT, and LINE1 was analyzed by pyrosequencing and compared with results from the cell lines RT4, T24, HeLa, BEAS-2B, and HepG2. Finally, short tandem repeat (STR) profiling was applied.Results:All tested UROtsa cell line stocks lacked large T-antigen. STR analysis unequivocally identified our main UROtsa stock as the bladder cancer cell line T24, which was different from two authentic UROtsa stocks that served as controls. Analysis of DNA methylation patterns and RNA expression confirmed their differences. Methylation pattern and mRNA expression of the contaminating T24 cell line showed moderate changes even after long-term culture of up to 56 weeks, whereas miRNAs and chromosome numbers varied markedly.Conclusions:It is important to check the identity of cell lines, especially those that are not distributed by major cell banks. However, for some cell lines STR profiles are not available. Therefore, new cell lines should either be submitted to cell banks or at least their STR profile determined and published as part of their initial characterization. Our results should help to improve the identification of UROtsa and other cells on different molecular levels and provide information on the use of urothelial cells for long-term experiments. © 2013 Johnen et al. OA gold
Databáze: OpenAIRE
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