FACS Enrichment of Total Interstitial Cells and Fibroblasts from Adult Mouse Ventricles
Nona Farbehi,
Vaibhao Janbandhu,
Robert Nordon,
Richard Harvey
Affiliations
Nona Farbehi
Victor Chang Cardiac Research Institute, Darlinghurst, Sydney, 2010, AustraliaGarvan Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Darlinghurst, Sydney, 2010, Australia
Vaibhao Janbandhu
Victor Chang Cardiac Research Institute, Darlinghurst, Sydney, 2010, AustraliaSt. Vincent's Clinical School, UNSW Sydney, Kensington, 2052, Australia
Robert Nordon
Graduate School of Biomedical Engineering, UNSW Sydney, Kensington, 2052, Australia
Richard Harvey
Victor Chang Cardiac Research Institute, Darlinghurst, Sydney, 2010, AustraliaSt. Vincent's Clinical School, UNSW Sydney, Kensington, 2052, Australia, School of Biotechnology and Biomolecular Science, UNSW Sydney, Kensington, 2052, Australia
Besides cardiomyocytes, the heart contains numerous interstitial cell types, including cardiac fibroblasts, endothelial cells, immune (myeloid and lymphoid) cells, and mural cells (pericytes and vascular smooth muscle cells), which play key roles in heart repair, regeneration, and disease. We recently published a comprehensive map of cardiac stromal cell heterogeneity and flux in healthy and infarcted hearts using single-cell RNA sequencing (scRNA-seq) (Farbehi et al., 2019). Here, we describe the FACS (Fluorescent Activated Cell Sorting)-based method used in that study for isolation of two cardiac cell fractions from adult mouse ventricles: the total interstitial cell population (TIP; non-cardiomyocytes) and enriched (Pdgfra-GFP+) cardiac fibroblasts.
