Abstract
The micronucleus (MN) assay, applied in different surrogate tissues, is one of the best validated cytogenetic techniques for evaluating chromosomal damage in humans. The cytokinesis-block micronucleus cytome assay (CBMNcyt) in peripheral blood lymphocytes is the most frequent method in biomonitoring human populations to evaluate exposure to genotoxic agents, micronutrient deficiency, or excess and genetic instability. Furthermore recent scientific evidence suggests an association between an increased MN frequency in lymphocytes and risk of cancer and other age-related degenerative diseases. The micronucleus cytome assay applied in buccal exfoliated cells (BMNCyt) provides a complementary method for measuring DNA damage and cytotoxic effects in an easily accessible tissue not requiring in vitro culture. The protocol for CBMNcyt described here refers to the use of ex vivo whole blood involving 72 h of culture with the block of cytokinesis at 44 h. BMNCyt protocol reports the established method for sample processing, slide preparation, and scoring.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Fenech M, Morley AA (1985) Measurement of micronuclei in lymphocytes. Mutat Res 147:29–36
Fenech M (2000) Mathematical model of the in vitro micronucleus assay predicts false negative results if micronuclei are not specifically scored in binucleated cells or in cells that have completed one nuclear division. Mutagenesis 15:329–336
Fenech M (2006) Cytokinesis-block micronucleus assay evolves into a “cytome” assay of chromosomal instability, mitotic dysfunction and cell death. Mutat Res 600:58–66
Fenech M (2001) The role of folic acid and vitamin B12 in genomic stability of human cells. Mutat Res 475:57–67
Fenech M, Aitken C, Rinaldi J (1998) Folate, vitamin B12, homocysteine status and DNA damage in young Australian adults. Carcinogenesis 19:1163–1171
Thomas P, Wu J, Dhillon V, Fenech M (2011) Effect of dietary intervention on human micronucleus frequency in lymphocytes and buccal cells. Mutagenesis 26(1):69–76
Dhillon VS, Thomas P, Iarmarcovai G et al (2011) Genetic polymorphisms of genes involved in DNA repair and metabolism influence micronucleus frequencies in human peripheral blood lymphocytes. Mutagenesis 26:33–42
El-Zein R, Vral A, Etzel CJ (2011) Cytokinesis-blocked micronucleus assay and cancer risk assessment. Mutagenesis 26(1):101–106
Bonassi S, Znaor A, Ceppi M et al (2007) An increased micronucleus frequency in peripheral blood lymphocytes predicts the risk of cancer in humans. Carcinogenesis 28:625–631
Bonassi S, El-Zein R, Bolognesi C et al (2011) Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies. Mutagenesis 26(1):93–100
Murgia E, Ballardin M, Bonassi S et al (2008) Validation of micronuclei frequency in peripheral blood lymphocytes as early cancer risk biomarker in a nested case-control study. Mutat Res 639:27–34
Migliore L, Coppedè F, Fenech M et al (2011) Association of micronucleus frequency with neurodegenerative diseases. Mutagenesis 26(1):85–92
Andreassi MG, Barale R, Iozzo P et al (2011) The association of micronucleus frequency with obesity, diabetes and cardiovascular disease. Mutagenesis 26(1):77–83
Holland N, Bolognesi C, Kirsch-Volders M et al (2008) The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage: the HUMN project perspective on current status and knowledge gaps. Mutat Res 659:93–108
Kirsch-Volders M, Mateuca RA, Roelants M et al (2006) The effects of GSTM1 and GSTT1 polymorphisms on micronucleus frequencies in human lymphocytes in vivo. Cancer Epidemiol Biomarkers Prev 15:1038–1042
Bonassi S, Neri M, Lando C et al (2003) HUMN collaborative group effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the Human MicroNucleus project. Mutat Res 543:155–166
Fenech M, Bonassi S (2011) The effect of age, gender, diet and lifestyle on DNA damage measured using micronucleus frequency in human peripheral blood lymphocytes. Mutagenesis 26(1):43–49
Fenech M, Holland N, Chang WP et al (1999) The HUman MicroNucleus Project: an international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans. Mutat Res 428:271–283
Fenech M, Chang WP, Kirsch-Volders M et al (2003) HUman MicronNucleus project. HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. Mutat Res 534:65–75
Bonassi S, Fenech M, Lando C et al (2001) HUman MicroNucleus project: international database comparison for results with the cytokinesis-block micronucleus assay in human lymphocytes—I. Effect of laboratory protocol, scoring criteria, and host factors on the frequency of micronuclei. Environ Mol Mutagen 37(1):31–45
Fenech M, Bonassi S, Turner J et al (2003) HUman MicroNucleus project, intra- and inter-laboratory variation in the scoring of micronuclei and nucleoplasmic bridges in binucleated human lymphocytes. Results of an international slide–scoring exercise by the HUMN project. Mutat Res 534:45–64
Fenech M, Bolognesi C, Kirsch-Volders M et al (2007) Harmonisation of the micronucleus assay in human buccal cells: a human micronucleus (HUMN) project (www.humn.org) initiative commencing in 2007. Mutagenesis 22:3–4
Bonassi S, Biasotti B, Kirsch-Volders M et al (2009) State of the art survey of the buccal micronucleus assay: a first stage in the HUMN(XL) project initiative. Mutagenesis 24:295–302
Fenech M (2007) Cytokinesis-block micronucleus cytome assay. Nat Protoc 2(5):1084–1104
Thomas P, Holland N, Bolognesi C et al (2009) Buccal micronucleus cytome assay. Nat Protoc 4:825–837
Lee TK, O'Brien K, Eaves GS et al (1999) Effect of blood storage on radiation-induced micronuclei in human lymphocytes. Mutat Res 444:201–206
Decordier I, Papine A, Vande LK et al (2011) Automated image analysis of micronuclei by IMSTAR for biomonitoring. Mutagenesis 26:163–168
Darzynkiewicz Z, Smolewski P, Holden E et al (2011) Laser scanning cytometry for automation of the micronucleus assay. Mutagenesis 26:153–161
Varga D, Johannes T, Jainta S et al (2004) An automated scoring procedure for the micronucleus test by image analysis. Mutagenesis 19:391–397
Bolognesi C, Balia C, Roggieri P et al (2011) Micronucleus test for radiation biodosimetry in mass casualty events: evaluation of visual and automated scoring. Rad Measurements 46:169–175
Bull CF, Mayrhofer G, Zeegers D et al (2012) Folate deficiency is associated with the formation of complex nuclear anomalies in the cytokinesis-block micronucleus cytome assay. Environ Mol Mutagen 53:311–323
Nersesyan A, Kundi M, Atefie K et al (2006) Effect of staining procedures on the results of micronucleus assays with exfoliated oral mucosa cells. Cancer Epidemiol Biomarkers Prev 15:1835–1840
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Bolognesi, C., Fenech, M. (2013). Micronucleus Assay in Human Cells: Lymphocytes and Buccal Cells. In: Dhawan, A., Bajpayee, M. (eds) Genotoxicity Assessment. Methods in Molecular Biology, vol 1044. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-529-3_10
Download citation
DOI: https://doi.org/10.1007/978-1-62703-529-3_10
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-528-6
Online ISBN: 978-1-62703-529-3
eBook Packages: Springer Protocols