Using single-cell RNA-Seq for unbiased analysis of developmental hierarchies (scRNA-Seq of BM lineage-negative Sca1+ CD117+ CD150+CD48-cells)

Hematopoietic stem cell metabolic requirements change with their cell cycle activity. However, the underlying role of mitochondria remain ill-defined. The goal of our study is to investigate how hematopoietic stem cells use mitochondria during cell division to control their fate and self-renewal activity. We found that after mitochondrial activation with replication, HSC irreversibly remodel the mitochondrial network and this network is not repaired after HSC re-entry into quiescence. HSC keep and accumulate dysfunctional mitochondria through asymmetric segregation during active division. Single cell RNA seq and bioinformatic analysis are used to characterize hematopoietic stem cell functions before and after replicative stress to uncover novel key drivers that limit hematopoietic stem cell self-renewal after replicative stress. Overall design: Single cell RNA seq hematopoietic stem cell populations; all 305 samples represent the 'scRNA-Seq of BM lineage-negative Sca1+ CD117+ CD150+CD48-cells'