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. LSK HSPCs were harvested isolated and sorted from flushed and crushed murine tibias and femurs. LSK HSPCs (WT, Fli-1ROSAΔ , Fli-1ROSAΔN1-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.