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Cytokinesis-Block Micronucleus Cytome Assay in Lymphocytes

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DNA Damage Detection In Situ, Ex Vivo, and In Vivo

Part of the book series: Methods in Molecular Biology ((MIMB,volume 682))

Abstract

The cytokinesis-block micronucleus cytome (CBMN cyt) assay is a new and comprehensive technique for measuring DNA damage, cytostasis, and cytotoxicity in different tissue types, including lymphocytes. DNA damage events are scored specifically in once-divided binucleated cells. These events 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. The assay has been applied to the biomonitoring of in vivo exposure to genotoxins, in vitro genotoxicity testing and in diverse research fields, such as nutrigenomics and pharmacogenomics. It has also been shown to be important as a predictor of normal tissue and tumor radiation sensitivity and cancer risk. This protocol also describes the current established methods for culturing lymphocytes, slide preparation, cellular and nuclear staining, scoring criteria, data recording, and analyses.

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Author information

Authors and Affiliations

  1. CSIRO Human Nutrition, Adelaide, SA, Australia

    Philip Thomas & Michael Fenech

Authors
  1. Philip Thomas
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  2. Michael Fenech
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Editor information

Editors and Affiliations

  1. Baylor College of Medicine, Depts. Neurosurgery and Molecular & Cell, University of Houston, Holcombe Blvd. 2002, Houston, 77030, Texas, USA

    Vladimir V. Didenko

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Thomas, P., Fenech, M. (2011). Cytokinesis-Block Micronucleus Cytome Assay in Lymphocytes. In: Didenko, V. (eds) DNA Damage Detection In Situ, Ex Vivo, and In Vivo. Methods in Molecular Biology, vol 682. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-409-8_16

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  • DOI: https://doi.org/10.1007/978-1-60327-409-8_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-408-1

  • Online ISBN: 978-1-60327-409-8

  • eBook Packages: Springer Protocols

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