GSE157821
Single cell RNA-seq of mouse HSC treated with iron chelator deferoxamine (DFO)
We characterized longer-term effects of iron homeostasis activation on hematopoietic stem cells by characterizing transcriptomic chagnes at the singe cell level 48hrs after iron chelatior treatement (vehicle treatemnt as controls). Overall design: 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 8-weeks. Lineage-depleted bone marrow cells from 8 mice were pooled and prospective HSC (Lin-Sca-1+c-Kit+CD150+CD48-) were sorted on MoFlo Astrios (Beckman Coulter). Thereafter, FACS-sorted HSC were exposed to iron chelator (DFO, Sigma) or vehicle (water) for 48 hours. Treated cells were collected and viability of >95% was determined by trypan blue exclusion. Twenty thousand (20,000) cells from each treatment group were then subjected to 10x Genomics platform for library preparation. Following quality control assessment with Agilent 2100 Bioanalyzer, approximately 50ng libraries were sequenced on BGISEQ-500 platform.