Best practices for authenticating cell lines
Abstract
Experiments using cell cultures are only valid to the extent that the cell culture is a true model system for the biological system being investigated. To assure that a cell line is and remains an appropriate biological model, its identity, purity, ploidy, and phenotype must be maintained. These characteristics comprise and determine the authenticity of a cell line. Routine monitoring of the cell line through microscopic examination of morphology can help to determine authenticity, as can the determination of phenotypic status. Assays designed to confirm cell identity and ploidy and freedom from cross-contaminating cell types may need to be performed at certain times, as such information may not be obtained through morphologic and phenotypic examinations alone. The best practices associated with establishing cell line authenticity are described in this article.
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Conventional karyotyping has been reported to detect contamination when present in as little as 1% of a culture.
“Conventional karyotyping has been reported to detect contamination when present in as little as 1% of a culture [31,97].”
From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [97]
DNA fingerprinting can detect contamination between human cell lines when it composes at least 5% of a culture.
“DNA Fingerprinting may have difficulty discriminating between closely-related cells, but it can detect contamination between human cell lines when it composes ≥ 5% of a culture [31,97].”
From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [97]
Isoenzyme and allozyme analysis can generally detect contamination affecting at least 5-10% of the culture population.
“The electrophoretic mobilities of chosen enzymes from a given cell are used to generate a unique gel banding pattern that can generally detect contamination affecting ≥ 5-10% of the culture population [24,31,78,97].”
From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [97]
SNP surveys detect contamination at or above 3%, and the cytochrome c oxidase I gene reveals contamination at or above 1%.
“Alternatives surveying single nucleotide polymorphisms (”
From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [97]
Cites
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“Technologies with the capability of detecting interspecies mixed cultures include karyotyping (determination of modal chromosome number) and multiplex DNA barcode assays designed to detect the most common species of cultured cells (e.g., Cooper et al. 2007).”
BackgroundMethodKleensang, A., Vantangoli, M. M., Odwin-DaCosta, S., Andersen, M. E., Boekelheide, K., Bouhifd, M., Fornace, A. J., Li, H.-H., Livi, C. B., Madnick, S., et al. (2016) · Scientific Reports · doi:10.1038/srep28994 · Cited by 111 · cited 1× · see citing works
“Cell phenotype can change as a result of changes in culture conditions, or as a result of genetic heterogeneity (Frattini et al. 2015; Kleensang et al. 2016).”
BackgroundHuman Biosample Authentication Using the High-Throughput, Cost-Effective SNPtraceTM System
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“…include short tandem repeat (STR) profiling for human cells (Masters et al. 2001; Reid et al. 2013), mouse cells (Almeida et al. 2014), dog cells (Berger et al. 2014), and certain primate cells (Almeida et al. 2011); and singlenucleotide repeat (SNP) profiling for human (Liang-Chu et al. 2015)…”
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The history, landscape, and outlook of human cell line authentication and security
Harbut, E., Makris, Y., Pertsemlidis, A., & Bleris, L. (2024) · SLAS Discovery · doi:10.1016/j.slasd.2024.100194 · Cited by 8 · see citing works
- Authors
- Nims, R. W., & Reid, Y.
- Affiliations
- American Type Culture Collection (US)
- Corresponding author
- Nims, Raymond W.
- ORCID
- Nims, Raymond W.
- Venue
- In Vitro Cellular & Developmental Biology - Animal
- Published
- 2017-12
- Volume
- 53
- Issue
- 10
- Pages
- 880-887
- ISSN
- 1071-2690 (ISSN-L)
- 1543-706X
- OpenAlex
- W2773448995
- Cited by
- 21
- References
- 5
- Citations by year
- 2024: 2 · 2023: 1 · 2022: 4 · 2021: 4 · 2020: 3 · 2019: 1 · 2018: 6
- FWCI
- 1.12
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- In Vitro Cellular & Developmental Biology - Animal (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Immunology and Microbiology › Microbiology › Microbial infections and disease research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- cell line authentication, ploidy, cross-contamination, cell morphology, cell phenotype, cell culture quality control, best practices
- MeSH terms
- Animals, Cell Line, Humans, Molecular Biology, Phenotype, Ploidies, Cell Culture Techniques, Cytogenetic Analysis
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