Microarray gene expression profiling of mouse HSC treated with iron chelator deferoxamine (DFO)

We delineted the role for the intracellular labile iron pool in coordinating a cascade of molecular events which reinforce hematopoietic stem cell (HSC) identity by charaterizing and comparing the transciptome of HSC treated for 16hrs with an iron chlator (DFO, 10uM) vs. mock controls. For isolation of mouse HSC, bone marrow cells were isolated from tibiae, femurs, pelvic bones and vertebrae of wildtype C57/B6 mice at the age of 6 to 10-weeks. Low-density bone marrow mononuclear cells (BMMNC) were enriched by density gradient centrifugation using Histopague 1083 (Sigma-Aldrich, MO). BMMNC from 4 mice were then subjected to prospective isolation of HSC (Lin−Sca-1+c-Kit+CD150+CD48−) on a MoFlo Astrios platform (Beckman Coulter). Thereafter, FACS-sorted HSC from each biological replicate were exposed to the treatment of iron chelator (DFO, Sigma) or vehicle (water) for 16 hours.