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The Liverbeads as a tool for the comet assay
Laurence Vian1, Ayda Yusuf, Claire Guyomard
1Laboratoire de Toxicologie, Faculté de Pharmacie, BP 14491, 34093 Montpellier Cedex 5, France. lvian@pharma.univ-montp1.fr
Mutation Research
|August 6, 2002
Summary
Liverbeads, a novel cell system, effectively activate promutagens into DNA-reactive metabolites. This makes them a suitable tool for detecting genotoxicity using the comet assay.
Area of Science:
- Hepatocyte research
- Genotoxicity testing
- Biotechnology
Background:
- Cryopreserved hepatocytes entrapped in alginate (Liverbeads) offer a stable cell model.
- The comet assay is a sensitive method for detecting DNA strand breaks in individual cells.
Purpose of the Study:
- To evaluate Liverbeads' suitability for the comet assay.
- To assess Liverbeads' capacity to metabolize promutagens into DNA-reactive species.
Main Methods:
- Investigated experimental conditions including treatment time, cell density, and bead viability.
- Assessed the metabolic activation of promutagens (cyclophosphamide, benzo[a]pyrene, etc.) by Liverbeads.
- Utilized the alkaline comet assay with tail extent moment (TEM) for data analysis.
Main Results:
- Human and rat Liverbeads showed significant positive results after 12h incubation.
- Liverbeads demonstrated the capacity to metabolize xenobiotics, unlike HeLa S3 cells.
- The comet assay with Liverbeads detected DNA damage induced by promutagens.
Conclusions:
- Liverbeads maintain specific metabolic functions, making them a suitable tool for genotoxicity detection.
- Liverbeads are easy to handle and effective in the comet assay for identifying promutagens.
- This system enhances the detection of genotoxic compounds through metabolic activation.