Single cell RNA sequencing to identify the PDGFR-β expressing lung cells

Purpose:Single cell RNA sequencing to identify PDGFR-beta expressing cells in the normal adult lung Methods: Adult Pdgfrb-CreERT2, ROSA26R(Zs/+) mice were induced with tamoxifen (1mg/day for 5 days) and rested for 5 days. Lungs were then harvested and prepared to obtain single cell suspension. Live Zs+ and Zs- cells were isolated by FACS and hash-tagged. Cells were then pooled together in PBS with 0.04% bovine serum albumin and subjected to library preparation and DropSeq. scRNA-seq data was processed with Cell Ranger v 3.1.0. Reads were aligned to a modified version of the mouse transcriptome mm10, including the sequence for the gene ZsGreen1. The top cell barcodes selected by Cell Ranger were utilized for downstream analysis. Results: In lung mesenchymal cells, populations of fibroblasts and pericytes express Pdgfr-beta. Lung PDGFR-beta+ cells (Zs+ cells isolated from tamoxifen-induced Pdgfrb-CreERT2, ROSA26R(Zs/+)mice) and PDGFR-beta- cells (Zs- cells) were isolated by FACS and subjected to 10X Genomics single cell 3’ RNA-seq libraries construction and sequencing. Both PDGFR-beta+ and PDGFR-beta- were included in this sample. Pdgfrb-beta status was identified based on the expression of Zs transcript (and Pdgfrb transcript) in the data.