Single-cell isolation from mouse heart

Samples were obtained from Left ventricular at indicated time points. Single cell suspensions were prepared as described previously, with some modifications (20 (link)). Briefly, hearts were cut into small pieces and digested with 0.3 mg/ml collagenase II (Invitrogen, USA), 0.3 mg/ml dispase II (Sigma, USA), DNase I (Biosharp, China) and 2.5 mM Cacl₂ (Mackin, china) in HBSS solution (Invitrogen, USA) for 45 min at 37°C with gentle agitation. After the digestion, primary cardiac cells were obtained using Percoll (Solarbio ^®) gradient separation and passed through 70-μm cell strainer. The obtained cells were washed with RPMI-1640 cell culture medium for further analysis.