Identification of cross-contaminated animal cells by PCR and isoenzyme analysis

Rajendran, R.; Nagarajan, T.; Sivakumar, V.; Senthilkumar, Rajappa; Obulapathi, B. Bala; Thiagarajan, D.; Srinivasan, V.A.

doi:10.1007/s10616-009-9245-5

Cited by 15 publications

(12 citation statements)

References 24 publications

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“…Cross-contamination of cell lines causes mischaracterization, which has been demonstrated in several studies. HeLa [16] and its derivatives HeLa S3, AV3 [17] and WISH contaminations are notorious problems and may be simply detected by either isoenzyme [18] or STR profiling [19]. Our findings showed evidence of 46.9% (60/128) cross-contamination was caused by HeLa, affecting 31 cell lines, which were purported to have been established from 10 types of tumor (colon, liver, breast, stomach, lung, nasopharynx, tougue, laryngeal, salivary gland and bronchia) and 3 types of normal tissue (lung, intestine and liver).…”

Section: Resultsmentioning

confidence: 99%

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines

et al. 2017

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In recent years, biological research involving human cell lines has been rapidly developing in China. However, some of the cell lines are not authenticated before use. Therefore, misidentified and/or cross-contaminated cell lines are unfortunately commonplace. In this study, we present a comprehensive investigation of cross-contamination and misidentification for a panel of 278 cell lines from 28 institutes in China by using short tandem repeat profiling method. By comparing the DNA profiles with the cell bank databases of ATCC and DSMZ, a total of 46.0% (128/278) cases with cross-contamination/misidentification were uncovered coming from 22 institutes. Notably, 73.2% (52 out of 71) of the cell lines established by the Chinese researchers were misidentified and accounted for 40.6% of total misidentification (52/128). Further, 67.3% (35/52) of the misidentified cell lines established in laboratories of China were HeLa cells or a possible hybrid of HeLa with another kind of cell line. Furthermore, the bile duct cancer cell line HCCC-9810 and degenerative lung cancer Calu-6 exhibited 88.9% match in the ATCC database (9-loci), indicating that they were from the same origin. However, when we used 21-loci to compare these two cell lines with the same algorithm, the percent match was only 48.2%, indicating that these two cell lines were different. The SNP profiles of HCCC-9810 and Calu-6 also revealed that they were different cell lines. 150 cell lines with unique profiles demonstrated a wide range of in vitro phenotypes. This panel of 150 genomically validated cancer cell lines represents a valuable resource for the cancer research community and will advance our understanding of the disease by providing a standard reference for cell lines that can be used for biological as well as preclinical studies.

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Section: Resultsmentioning

confidence: 99%

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines

et al. 2017

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“…Karyotype is a method used to identify specifi c chromosome markers, the genus and species of the animal, to analyze the characteristic chromosome number of each species and also determine common aneuploid in cell lines (Markovic & Markovic 1998;Wenger et al 2005, Ramya 2009Dittmar et al 2010;Leandro & Cruz 2012). It's a method that takes more time and depends on an expert ability.…”

Section: Introductionmentioning

confidence: 99%

Isoenzyme Electrophoresis and Karyotype Techniques as an Alternative for Cell Line Identity Confirmation

Machado

Miranda

Gonçalves

et al. 2014

VR&R

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Th e use of cell culture in biomedical science has been constantly enhanced and the use of certifi ed and microrganismsfree cell lines is indispensable. Due to the value and the diffi culty of acquiring a commercially available kit (Authentikit TM system) for our routine in addition with the purpose of monitoring and ensuring quality of provided cell lines, and also contributing with cell characterization, the Núcleo de Cultura de Células do Instituto Adolfo Lutz (NCC-AL) padronized two techniques to identify the animal species from which the cells were originated: isoenzyme electrophoresis and karyotype. Cell extracts from 14 diff erent cell lines were run under horizontal electrophoresis in agarose gel and lactate dehydrogenase isoenzyme profi le was revealed using tetrazolium salt. Th e number of bands and the migration distances between bands of cell lines were similar to the literature, but small adjustments were needed to assist in the visualization of the bands that were not visible. Cell fl asks for karyotype were incubated with colcemid solution and modal values of each cell line were calculated. Comparing all the results obtained with those reported in the literature, we conclude that both methods are promising and will allow better control of the services provided by the laboratory, not to mention guaranteeing authenticated cell lines for research and diagnostics.

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“…RNase free water was used as negative control. Arrows indicate the DNA molecular weight marker of 500 bp (Hyperladder IV; Bioline) sensitivity or ability to detect contamination (Liu et al 2003;Ono et al 2007;Ramya et al 2009;Higgins et al 2010;Parodi et al 2002). Inexpensive assays for the identification and authentication of cell lines are therefore clearly necessary and should be relatively easily developed using the vast DNA databases as is demonstrated here.…”

Section: Discussionmentioning

confidence: 99%

Species identification and authentification of human and rodent cell cultures using polymerase chain reaction analysis of vomeronasal receptor genes

Holder

Cooper

2011

Cytotechnology

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Cell culture and the use of cell lines are routinely used in basic scientific research. It is therefore imperative for researchers to ensure the origin of the cell lines used and that they are routinely re-analysed for contamination and misidentification. Inter-species contamination is relatively frequent, and the most commonly used cell lines are of human, mouse and rat derivation. We have developed simple species specific primer assays based on genomic sequence differences in vomeronasal receptor gene family members to discriminate between human, mouse and rat DNA using standard agarose gel electrophoresis. Furthermore, these PCR assays are able to identify the species composition within an inter-species mixed population. This approach therefore provides a valuable tool to enable a rapid, simple and relatively inexpensive determination of the authentication and contamination of cell cultures.

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Identification of cross-contaminated animal cells by PCR and isoenzyme analysis

Cited by 15 publications

References 24 publications

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines

Isoenzyme Electrophoresis and Karyotype Techniques as an Alternative for Cell Line Identity Confirmation

Species identification and authentification of human and rodent cell cultures using polymerase chain reaction analysis of vomeronasal receptor genes

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