Analysis of metabolic enzyme expression reveals a unique progenitor cell with mast cell and erythrocyte potential

The purpose of this study was to take an unbiased look at the progenitor cell subsets that are present in the bone marrow of naïve mice. By using single cell RNA-seq we hoped to identify marker genes of interest that could then be tracked by the generation new reporter animals. First, CD45+ CD3- Ly6G- cells were sort-purified from the bone marrow of 3 naïve C57BL/6 mice and pooled. This was the input for the (Bone marrow-resident progenitor cells) sample. Second, CD45+ Car1-GFP-expressing cells were sort-purified from the bone marrow of 3 naïve Car1-GFP reported mouse on a C57BL/6 background and pooled. This was the input for the (Car1-GFP+ bone marrow cells) sample. 10,000 sort-purified cells were then processed using the 10X Genomics ChromiumTM Controller. Cell suspensions were loaded onto the Chromium Single Cell A Chip for cell lysis and barcoding. RNA from individual cells was reverse transcribed and sequencing libraries prepared using the ChromiumTM Single Cell 3’ Library Kit v2 following the manufacturers protocol. Samples were sequenced using an Illumina NextSeq 550 with standard 10X Genomics Configuration (26bpx98bp).