ABSTRACT: Using single-cell RNA-Seq for unbiased analysis of developmental hierarchies (single cell RNA Seq of bone marrow lineage-negative CD117+ CD34+ cells)
Project description:Bulk transcriptomes of granulocyte-macrophage progenitor cells isolated from bone marrow of Rnaseh2bfl/fl/ Vav-Cre and Cre-negative ctrl mice; The sorted bone marrow cells had the following immunophenotype: lineage- CD201 (EPCR)- CD117+ CD16/32+ CD34+
Project description:In this study we performed single-cell RNA sequencing on 7000 negatively enriched fresh NK and progenitor cells from one healthy bone marrow donor, that were pre-labeled with oligonucleotide-conjugated antibodies against CD34, CD117, CD56 and CXCR6. A new CD56dimGzmK+ subset was identified that shared features with CD56bright and CD56dimGzmK- NK cells based on transcriptome, phenotype (NKG2AhighCD16lowKLRG1highTIGIThigh) and functional analysis in bone marrow and blood, supportive for an intermediate subset. Pseudotime analysis positioned CD56bright, CD56dimGzmK+ and CD56dimGzmK- cells in one differentiation trajectory, while ltNK cells were separated in a distinct lineage. Integrative analysis with bone marrow cells from the human cell atlas did not demonstrate a developmental connection between CD34+ progenitor and NK cells, implicating that bone marrow is not a site of active NK cell development.
