Open AccessIn vitro erythropoiesis from bone marrow-derived progenitors provides a physiological assay for toxic and mutagenic compounds
Author(s) -
Joe Shuga,
J. Zhang,
Leona D. Samson,
Harvey F. Lodish,
Linda G. Griffith
Publication year - 2007
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0701829104
Subject(s) - erythropoiesis , genotoxicity , haematopoiesis , bone marrow , biology , micronucleus test , in vitro , in vivo , dna damage , erythropoietin , micronucleus , progenitor cell , microbiology and biotechnology , in vitro toxicology , clonogenic assay , cancer research , immunology , chemistry , stem cell , dna , biochemistry , genetics , toxicity , medicine , anemia , organic chemistry
The goal of this study was to create anin vitro cell culture system that captures essential features of thein 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 thisin 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 exposurein vivo compared with WT BM and that this phenotypic response was reflected in erythropoietic cultures. These findings suggest that thisin 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.