Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

Mutation Research
Steven M BryceStephen D Dertinger

Abstract

An international, multi-lab trial was conducted to evaluate a flow cytometry-based method for scoring micronuclei in mouse lymphoma L5178Y cells [S.L. Avlasevich, S.M. Bryce, S.E. Cairns, S.D. Dertinger, In vitro micronucleus scoring by flow cytometry: differential staining of micronuclei versus apoptotic and necrotic chromatin enhances assay reliability, Environ. Mol. Mutagen. 47 (2006) 56-66]. A reference laboratory investigated the potential of six chemicals to induce micronuclei -- the genotoxicants mitomycin C (MMC), etoposide (ETOPO), and vinblastine (VB), and the non-genotoxicants sucrose (SUC), staurosporine (STS), and dexamethasone (DEX). The latter two non-genotoxicants were selected as extreme challenges to the assay because of their potent apoptogenic activity. Three collaborating laboratories were supplied with prototype In Vitro MicroFlow kits, and each was assigned one genotoxicant and one non-genotoxicant. Cells were treated continuously for 24h over a range of concentrations up to 5 mg/ml, or overtly cytotoxic concentrations. Micronuclei were scored via standard microscopy and flow cytometry. In addition to enumerating micronucleus frequencies, a cytotoxicity measurement that is simultaneously acquired with the...Continue Reading

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Citations

Jan 1, 2014·Advanced Drug Delivery Reviews·Adrian Roth, Thomas Singer
Feb 22, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Benjamin C MoellerJames A Swenberg
Jan 15, 2016·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
Dec 14, 2011·Journal of Hazardous Materials·Daniela Morais LemeMaria Aparecida Marin-Morales
Jul 14, 2009·Regulatory Toxicology and Pharmacology : RTP·Andrew W KnightRichard M Walmsley
Jul 25, 2009·Mutation Research·Gary Peter WattersCaroline Ann Austin
Sep 28, 2010·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
Nov 26, 2010·British Journal of Pharmacology·Richard Maurice Walmsley, Nicholas Billinton
Dec 2, 2009·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
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Jul 2, 2016·Environmental and Molecular Mutagenesis·Derek T BernackiStephen D Dertinger
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Oct 31, 2019·Archives of Toxicology·Shuaizhang Li, Menghang Xia
Jul 4, 2020·Toxicological Sciences : an Official Journal of the Society of Toxicology·Inger BrandsmaGiel Hendriks

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis