title: In vitro erythropoiesis from bone marrow-derived progenitors provides a physiological assay for toxic and mutagenic compounds.
abstract: The goal of this study was to create an in vitro cell culture system that captures essential features of the in vivo erythroid micronucleus (MN) genotoxicity assay, thus enabling increased throughput and controlled studies of the hematopoietic DNA damage response. We show that adult bone marrow (BM) cultures respond to erythropoietin, the principal hormone that stimulates erythropoiesis, with physiological erythropoietic proliferation, differentiation, and enucleation. We then show that this in vitro erythropoietic system clearly signals exposure to genotoxicants through erythroid MN formation. Furthermore, we determined that DNA repair-deficient (MGMT(-/-)) BM displayed sensitivity to genotoxic exposure in vivo compared with WT BM and that this phenotypic response was reflected in erythropoietic cultures. These findings suggest that this in vitro erythroid MN assay is capable of screening for genotoxicity on BM in a physiologically reflective manner. Finally, responses to genotoxicants during erythroid differentiation varied with exposure time, demonstrating that this system can be used to study the effect of DNA damage at specific developmental stages.
authors.affiliation: Department of Chemical Engineering, Whitehead Institute for Biomedical Research, Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: J
authors.initials: J
authors.lastname: Shuga
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: L D
authors.initials: LD
authors.lastname: Samson
authors.forename: H F
authors.initials: HF
authors.lastname: Lodish
authors.forename: L G
authors.initials: LG
authors.lastname: Griffith
mesh_terms: D000818:Animals; D015415:Biomarkers; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D003603:Cytotoxins / Q000633:toxicity*; D045643:DNA Repair Enzymes / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D004920:Erythropoiesis / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015162:Micronucleus Tests; D016296:Mutagenesis; D009152:Mutagenicity Tests; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D003603:Cytotoxins; D045643:DNA Repair Enzymes