NextSeq 500 sequencing; Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury
Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury
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 to be filtered on 40μm strainers. Cells were then resuspended in red cell lysis buffer followed by dead cell removal and immunostaining for 15 minutes on ice with fluorophore-conjugated antibodies and followed by FACS sorting for mentioned markers. Immediately after cell sorting and counting, the cells were loaded on 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
Experimental Factor: injury
sham
Experimental Factor: time
7
Experimental Factor: fraction
total interstitial non-myocyte cell