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Cytokinesis-block micronucleus cytome assay

Journal

NATURE PROTOCOLS
Volume 2, Issue 5, Pages 1084-1104

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.77

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The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated ( BN) cells and include ( a) micronuclei ( MNi), a biomarker of chromosome breakage and/ or whole chromosome loss, ( b) nucleoplasmic bridges ( NPBs), a biomarker of DNA misrepair and/ or telomere end-fusions, and ( c) nuclear buds ( NBUDs), a biomarker of elimination of amplified DNA and/ or DNA repair complexes. Cytostatic effects are measured via the proportion of mono-, bi- and multinucleated cells and cytotoxicity via necrotic and/ or apoptotic cell ratios. Further information regarding mechanisms leading to MNi, NPBs and NBUDs formation is obtained using centromere and/ or telomere probes. The assay is being applied successfully for biomonitoring of in vivo genotoxin exposure, in vitro genotoxicity testing and in diverse research fields such as nutrigenomics and pharmacogenomics as well as a predictor of normal tissue and tumor radiation sensitivity and cancer risk. The procedure can take up to 5 days to complete.

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