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The in vitro MN assay in 2011: origin and fate, biological significance, protocols, high throughput methodologies and toxicological relevance

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Abstract

Micronuclei (MN) are small, extranuclear bodies that arise in dividing cells from acentric chromosome/chromatid fragments or whole chromosomes/chromatids lagging behind in anaphase and are not included in the daughter nuclei at telophase. The mechanisms of MN formation are well understood; their possible postmitotic fate is less evident. The MN assay allows detection of both aneugens and clastogens, shows simplicity of scoring, is widely applicable in different cell types, is internationally validated, has potential for automation and is predictive for cancer. The cytokinesis-block micronucleus assay (CBMN) allows assessment of nucleoplasmic bridges, nuclear buds, cell division inhibition, necrosis and apoptosis and in combination with FISH using centromeric probes, the mechanistic origin of the MN. Therefore, the CBMN test can be considered as a “cytome” assay covering chromosome instability, mitotic dysfunction, cell proliferation and cell death. The toxicological relevance of the MN test is strong: it covers several endpoints, its sensitivity is high, its predictivity for in vivo genotoxicity requires adequate selection of cell lines, its statistical power is increased by the recently available high throughput methodologies, it might become a possible candidate for replacing in vivo testing, it allows good extrapolation for potential limits of exposure or thresholds and it is traceable in experimental in vitro and in vivo systems. Implementation of in vitro MN assays in the test battery for hazard and risk assessment of potential mutagens/carcinogens is therefore fully justified.

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Acknowledgments

This work was supported by the EU research programmes “The Detection and Hazard Evaluation of Aneugenic Chemicals” (ENV4-CT97-0471), “Protection of the European Population from Aneugenic Chemicals (PEPFAC)” (QLK4-CT-2000-00058), ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5: “Food Quality and Safety” (Contract no. 513943) and by the EU Integrated Project NewGeneris (“Newborns and Genotoxic exposure risk”)’, 6th Framework Programme, Priority 5: Food Quality and Safety (Contract no. FOOD-CT-2005-016320). NewGeneris is the acronym of the project, the Belgian Science Policy [contractnumber SD/HE/02B]; OZR-VUB and the European Union small ormedium-scale focused research project ENPRA (Risk Assessment of Engineered Nanoparticles) (Contract no. NMP-2008-1.3-2).

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  1. Laboratorium voor Cellulaire Genetica, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium

    Micheline Kirsch-Volders, Gina Plas, Laetitia Gonzalez, Kim Vande Loock & Ilse Decordier

  2. Novartis Institutes for Biomedical Research, Translational Sciences, Preclinical Safety, Genetic Toxicology and Safety Pharmacology, MUT-2881.5.38, 4002, Basel, Switzerland

    Azeddine Elhajouji & Magdalena Lukamowicz

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  1. Micheline Kirsch-Volders
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Correspondence to Ilse Decordier.

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Kirsch-Volders, M., Plas, G., Elhajouji, A. et al. The in vitro MN assay in 2011: origin and fate, biological significance, protocols, high throughput methodologies and toxicological relevance. Arch Toxicol 85, 873–899 (2011). https://doi.org/10.1007/s00204-011-0691-4

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