Flow-cytometric analysis of in vitro micronucleus formation: comparative studies with WIL2-NS human lymphoblastoid and L5178Y mouse lymphoma cell lines

Mutation Research
S LaingamA R Humpage

Abstract

The development of a flow cytometry-based micronucleus (FCMMN) assay for measuring the micronucleus (MN) frequency in cells following exposure to test chemicals has potential for improving reproducibility and reducing turn-around time when compared with the traditional microscopy-based micronucleus method. A major drawback of the FCMMN assay is that a false-positive interpretation could result from the presence of large numbers of apoptotic or necrotic bodies in the measured sample. Although several studies have reported ways in which the FCMMN assay could be improved using different staining techniques or electronic gating strategies, to date none of these protocols are suitable for use as a screening assay. To reduce the interference from apoptosis, performing the FCMMN assay with an apoptosis-resistant cell line may be an alternative approach. This study reports the use of p53-mutated cell lines to minimise the interference found in the FCMMN assay. Two commonly used cell lines (WIL2-NS and L5178Y) were investigated by comparison of (1) cytotoxicity and micronucleus induction in the FCMMN assay following treatment with model genotoxicants and (2) apoptotic responses after exposure to inducers of apoptosis. Both cell lines we...Continue Reading

References

Dec 13, 2000·Mutation Research·M Fenech
Aug 16, 2001·Critical Reviews in Toxicology·E J Calabrese, L A Baldwin
Aug 16, 2001·Critical Reviews in Toxicology·E J Calabrese
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Jan 29, 2003·Environmental and Molecular Mutagenesis·Sophie MeintièresDaniel Marzin
Jan 10, 2008·Mutation Research·Steven M BryceStephen D Dertinger

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Citations

Jul 25, 2009·Mutation Research·Gary Peter WattersCaroline Ann Austin
Dec 2, 2008·Mutation Research·Ana María SalazarPatricia Ostrosky-Wegman
Mar 10, 2012·Environmental and Molecular Mutagenesis·S LaingamA R Humpage
Aug 2, 2015·Mutation Research. Genetic Toxicology and Environmental Mutagenesis·James WhitwellVeronique Thybaud
Aug 28, 2020·Nanomaterials·Bwalya A WitikaRoderick B Walker
Dec 29, 2013·Environmental Science & Technology·Beate I EscherFrederic D L Leusch

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