Illumina NovaSeq 6000 sequencing; Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury and PDGF-AB or PBS delivery
Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury and PDGF-AB or PBS delivery
Tissue from ventricles of male adult mice was enzymatically digested into a single cell suspension and viable non-myocte cells were isolated using FACS sorting. Briefly, hearts were minced and incubated in collagenase type II (Worthington, USA) at 37°C before being filtered on 40μm strainers. Cells were then resuspended in red cell lysis buffer followed by dead cell removal and live cells (DAPI-) were FACS sorted and counted to be loaded on to the 10x Chromium microfluidics chip (3-prime V2). Cells were loaded on the 10x Chromium system and single cell RNA-seq libraries were generated following the manufacturer's instructions. 10x Chromium 3-prime V2 protocol 10x Chromium 3-prime V2 protocol