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A novel micronucleus in vitro assay utilizing human hematopoietic stem cells
Stockholm Univ, Dept Mol Biosci, Wenner Gren Inst, SE-10691 Stockholm, Sweden..
Univ Paris 06, UMR CDR St Antoine Proliferat & Differentiat Cell, Paris, France.;INSERM, UMR Proliferat & Differentiat Cellules Souches S9, Paris, France.;Etab Francais Sang Ile France, Ivry, France..
Stockholm Univ, Dept Mol Biosci, Wenner Gren Inst, SE-10691 Stockholm, Sweden..
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2015 (English)In: Toxicology in Vitro, ISSN 0887-2333, E-ISSN 1879-3177, Vol. 29, no 7, 1897-1905 p.Article in journal (Refereed) Published
Abstract [en]

The induction of micronucleated reticulocytes in the bone marrow is a sensitive indicator of chromosomal damage. Therefore, the micronucleus assay in rodents is widely used in genotoxicity and carcinogenicity testing. A test system based on cultured human primary cells could potentially provide better prediction compared to animal tests, increasing patient safety while also implementing the 3Rs principle, i.e. replace, reduce and refine. Hereby, we describe the development of an in vitro micronucleus assay based on animal-free ex vivo culture of human red blood cells from hematopoietic stem cells. To validate the method, five clastogens with direct action, three clastogens requiring metabolic activation, four aneugenic and three non-genotoxic compounds have been tested. Also, different metabolic systems have been applied. Flow cytometry was used for detection and enumeration of micronuclei. Altogether, the results were in agreement with the published data and indicated that a sensitive and cost effective in vitro assay to assess genotoxicity with a potential to high-throughput screening has been developed. (C) 2015 The Authors. Published by Elsevier Ltd.

Place, publisher, year, edition, pages
2015. Vol. 29, no 7, 1897-1905 p.
Keyword [en]
Stem cells, Erythrocytes, Micronucleus test, Flow cytometry, Genotoxicity in vitro
National Category
Pharmacology and Toxicology
Identifiers
URN: urn:nbn:se:uu:diva-264618DOI: 10.1016/j.tiv.2015.07.018ISI: 000361263500063PubMedID: 26208286OAI: oai:DiVA.org:uu-264618DiVA: diva2:865095
Funder
Swedish Research Council
Available from: 2015-10-26 Created: 2015-10-15 Last updated: 2017-12-01Bibliographically approved

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