Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling (scRNAseq)

CD44+ and CD44- negative Alveolar type 2 cells (AT2) represent distinct populations in lung fibrosis. To charecterize these cells, we isolated CD44+/- AT2s from WT mice and from fibrotic Sftpc muant lungs and performd RNA sequencing no these populations Overall design: To enrich for EPCAM+ cells and mesenchymal cells , a single cell suspension was prepared by physical and enzymatic dissociation (Collagenase, DNase, Dispase) from Sftpc mutation firbosis lungs (n=2) and control lungs (n=2) followed by MACS sorting by LS columns with CD45+ and CD31 cell removal using CD45 and CD31 micobeads followed by EPCAM+ cell enrichment using EPCAM microbeads. CD45-/CD31-/EPCAM- cells were considered lung mesenchymal cells. EPCAM+ and mesenchymal cells were prepared in 50/50 concentration for sequencing.