Transcriptional Activation of Regenerative Haematopoiesis via Niche Sensing [scRNA-seq cocultured HSPCs]

Single cell RNA-seq analysis was performed on sorted population of LSK HSPCs with different genetic background and phenotype, after co-culture with vascular niche cells, to uncover differential transcriptional output following genetic manipulation. Overall design: LSK HSPCs were harvested isolated and sorted from flushed and crushed murine tibias and femurs. LSK HSPCs (WT, Fli-1ROSA? , Fli-1ROSA?N1-ICiOE, and N1-ICiOE) were introduced into co-culture with vascular niche ECs. 4-OHT was introduced into cultures to induce genetic manipulation. Cultures were harvested and LSK HSPCs were resorted and processed for 10X Chromium scRNA-seq analysis.