title: A novel micronucleus in vitro assay utilizing human hematopoietic stem cells.
abstract: 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.
authors.affiliation: Stockholm University, Department of Molecular Biosciences, The Wenner-Gren Institute, SE-106 91 Stockholm, Sweden. Electronic address: Natalia.Kotova@su.se.
authors.forename: N
authors.initials: N
authors.lastname: Kotova
authors.affiliation: UPMC Univ Paris 06, UMR_S938 CDR Saint-Antoine, Prolifération et Différentiation des Cellules Souches, Paris, France; INSERM, UMR_S938, Prolifération et Différentiation des Cellules Souches, Paris, France; Etablissement Francais du Sang Ile de France, Ivry-sur-Seine, France.
authors.forename: N
authors.initials: N
authors.lastname: Hebert
authors.affiliation: Stockholm University, Department of Molecular Biosciences, The Wenner-Gren Institute, SE-106 91 Stockholm, Sweden.
authors.forename: E-L
authors.initials: EL
authors.lastname: Härnwall
authors.affiliation: Stockholm University, Department of Molecular Biosciences, The Wenner-Gren Institute, SE-106 91 Stockholm, Sweden.
authors.forename: D
authors.initials: D
authors.lastname: Vare
authors.affiliation: UPMC Univ Paris 06, UMR_S938 CDR Saint-Antoine, Prolifération et Différentiation des Cellules Souches, Paris, France; INSERM, UMR_S938, Prolifération et Différentiation des Cellules Souches, Paris, France; Etablissement Francais du Sang Ile de France, Ivry-sur-Seine, France.
authors.forename: C
authors.initials: C
authors.lastname: Mazurier
authors.affiliation: UPMC Univ Paris 06, UMR_S938 CDR Saint-Antoine, Prolifération et Différentiation des Cellules Souches, Paris, France; INSERM, UMR_S938, Prolifération et Différentiation des Cellules Souches, Paris, France; Etablissement Francais du Sang Ile de France, Ivry-sur-Seine, France; AP-HP, Hôpital Saint-Antoine, Service d'Hématologie Biologique, Paris, France.
authors.forename: L
authors.initials: L
authors.lastname: Douay
authors.affiliation: Stockholm University, Department of Molecular Biosciences, The Wenner-Gren Institute, SE-106 91 Stockholm, Sweden.
authors.forename: D
authors.initials: D
authors.lastname: Jenssen
authors.affiliation: Dept. of Genetics and Pathology, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Grawé
mesh_terms: D002478:Cells, Cultured; D018920:Coculture Techniques; D004912:Erythrocytes / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D015162:Micronucleus Tests; D009153:Mutagens / Q000633:toxicity*
chemical_list: D009153:Mutagens
keywords: Erythrocytes; Flow cytometry; Genotoxicity in vitro; Micronucleus test; Stem cells