GSM6422866: WTMI_24H; Mus musculus; RNA-Seq
Each mouse was euthanized via neck dislocation and the heart was explanted and rinsed with ice-cold HBSS. Subsequently, the ventricles were cut in pieces and incubated at 37°C for 30 minutes for enzymatic digestion (using thermolysin and papain). After 30 minutes the tissue pieces were resuspended several times to dissociate cells from fibrous tissue. The suspension was filtered via 300um mesh to get rid of big fibrous tissues and then 40um mesh to filter out cardiomyocytes. The resulting non-cardiomyocytes (NCM) suspension was then incubated with Calcein-AM and DAPI and live cells were sorted using FACS (fluorescence activated cell sorting). The live NCM from 4 different mice hearts were pooled for each sample before performing single-cell RNAseq. 10x 3-RNA-seq