Project description:4 months old Scl-tta; H2B-GFP mice where chased with Doxycyclin for 18 months from and then sacrificed at 22 months of age. GFP+ and GFP- cells, or dormant and active HSCs were then sorted and sequenced.
Project description:Quiescent hematopoietic stem cells (HSCs) are typically dormant, and only a few quiescent HSCs are active. The relationship between “dormant” and “active” HSCs remains unresolved. Here we generated a G0 marker (G0M) mouse line that visualizes quiescent cells. G0M identifies a small population of active HSCs (G0Mlow), which are distinct from dormant HSCs (G0Mhigh), within the conventional quiescent HSC fraction. Single-cell RNA-Seq analyses showed that the gene expression profiles of these populations were nearly identical, yet differed in their Cdk4/6 activity. Furthermore high-throughput small molecule screening revealed that high concentrations of cytoplasmic calcium ([Ca2+]c) were linked to dormancy of HSCs. These findings indicate that G0M separates dormant and active adult HSCs, which are regulated by Cdk4/6 and [Ca2+]c. This G0M mouse line represents a useful resource for investigating physiologically important stem cell subpopulations.
Project description:Identification of differentially expressed genes in young (3 month old) versus aged (24 month old) mouse hematopoietic stem cells. Comparison of genes differentially expressed in hematopoietic stem cells of young mice with conditional deletion of mTOR within vascular endothelium.
Project description:Rare dormant hematopoietic stem cells (dHSCs) reside at the top of the blood hierarchy harboring the highest long-term reconstitution capacity. However, no markers exist to prospectively identify dHSCs and their molecular identity, as well as the mechanism leading to their activation remains poorly understood. Here, we present the global transcriptome of ex vivo isolated mouse multipotent hematopoietic stem cells (HSCs) and dHSCs at the single cell level.
Project description:Rare dormant hematopoietic stem cells (dHSCs) reside at the top of the blood hierarchy harboring the highest long-term reconstitution capacity. Here, we present the global transcriptome of ex vivo isolated mouse multipotent dormant hematopoietic stem cells (dHSC), active HSCs (aHSCs) and multipotent progenitor cells (MPP1) as revealed by next-generation sequencing (RNA-seq) at the population level.