Identification of cells in culture

Stulberg, Cyril S.; Peterson, Ward D.; Simpson, W. F.

doi:10.1002/ajh.2830010208

Cited by 35 publications

(11 citation statements)

References 18 publications

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“…Using species specific antigens, isoenzyme electrophoresis, and chromosome analysis, he reported a frequency of 14% for interspecies contamination and 25% for intraspecies contamination. He concluded that, overall, 30% of the cell lines examined were incorrectly designated (Stulberg et al, 1976). At that time, a term "allozyme genetic signatures" was introduced by O'Brien.…”

Section: Prior Experience With Cell Cross-contamination: Evolution Ofmentioning

confidence: 97%

Cell cross-contamination in cell cultures: The silent and neglected danger

Marković

Marković²

1998

In Vitro Cell.Dev.Biol.-Animal

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Cell cross-contamination in cell cultures is a common problem during cell culturing and use. Contamination invalidates research results, compromises the comparison of results between laboratories, reduces reproducibility required in industrial production of cell lines, and may lead to unusable therapeutic products. The problem can be solved by increasing the awareness of its seriousness and by introducing regular quality control of cell cross-contamination in every laboratory where cells are grown and used.

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Section: Prior Experience With Cell Cross-contamination: Evolution Ofmentioning

confidence: 97%

Cell cross-contamination in cell cultures: The silent and neglected danger

Marković

Marković²

1998

In Vitro Cell.Dev.Biol.-Animal

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“…Several methods had been established to monitor cell culture identity using immunological markers (Stulberg et al 1976;O'Toole et al 1983), isoenzyme analysis (Halton et al 1983;Steube et al 1995) and karyologic examination (Miller et al 1971;NelsonRees et al 1974). The development of DNA-fingerprinting techniques offered detection of individual human samples (Jeffreys et al 1985;Stacey et al 1992) and is now used as an international standard for authentication of human cell lines Masters et al 2001).…”

Section: Introductionmentioning

confidence: 98%

Identification and verification of rodent cell lines by polymerase chain reaction

2007

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Cell lines represent valuable tools for basic research and diagnostic applications as well as for the production of biological products such as antibodies or vaccines. For all cell culturists, a well-identified origin of their cell lines as well as the periodic re-examination of their identity should be a basic requirement. We established a simple polymerase chain reaction (PCR) to verify or identify rodent and human cell lines. Since mouse-, rat-, Chinese hamster- and Syrian hamster-derived cell lines represent the most frequently used rodent cell lines, our investigations were focused on these species. Our assay used oligonucleotide primers annealing to sequences within the beta-actin and the beta-globin gene and to repetitive DNA. Primers were designed mostly from intron sequences of the genes aiming to amplify only one specific DNA segment and thus enabling to exclude easily false DNA. More than 130 cells lines originating from the five species were analyzed in that study. Our PCR revealed specific profiles for all species investigated. No further methods like DNA sequencing or fragment length polymorphism analysis were needed to differentiate these species. The results introduce our PCR-assay as a rapid, specific and routinely feasible tool in order to identify or distinguish rodent cell lines from each other and from human cell lines.

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“…Several methods had been established to monitor cell culture identity using immunological marker (Stulberg et al 1976;O'Toole et al 1983), isoenzyme analysis (Halton et al 1983;Steube et al 1995) and karyological examination (Miller et al 1971;Nelson-Rees et al 1974). Speciesspecific antibodies are also available commercially and these have been used to confirm species of origin by indirect fluorescence or immunoperoxidase staining (Hay 2002).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2009

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Animal cell lines have become very popular substrates for the production of vaccines and biopharmaceuticals. Characterization of candidate production cell lines is central to ensure product safety and maintenance of consistency in the manufacture of biologicals. Nested PCR and isoenzyme analysis have been used widely to prove the identity and purity of various cell lines and primary cells individually and also after deliberate cross-contamination. The nested PCR based on the Cytochrome b (Cyt b) gene of mitochondrial DNA (Mt DNA) was found to be more sensitive than isoenzyme analysis in detecting low levels of contaminants (as low as 1%). Interestingly, competition between different co-cultured cell lines has shown in one case that cross-contamination need not always results in a mixed cell population. The nested PCR technique for the Cyt b gene described in this study appears to be a potential replacement for isoenzyme analysis and here we demonstrate the PCR method used is sensitive and reliable for cell line authentication in a simple, rapid and reliable format to help assure the authenticity of cell substrates for the production of safe vaccines and biopharmaceuticals.

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Identification of cells in culture

Cited by 35 publications

References 18 publications

Cell cross-contamination in cell cultures: The silent and neglected danger

Cell cross-contamination in cell cultures: The silent and neglected danger

Identification and verification of rodent cell lines by polymerase chain reaction

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

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