The Culture of Primary Neonatal Mouse Cardiomyocytes

The culture of primary cardiomyocytes was described previously (Ehler et al., 2013). Briefly, cells were cultured in complete Dulbecco's Modified Eagle's Medium (DMEM, HyClone, USA) with 4,500 mg/L glucose,4 mM L-glutamine, 110 mg/L sodium pyruvate, 1% (v/v) penicillin/streptomycin and 10% (v/v) fetal bovine serum (FBS, Biological Industries). Cells were then placed in an incubator containing 95% air and 5% CO₂ at 37°C. The culture medium was replaced every 2–3 days. For the in vitro study, primary cardiomyocytes were treated with low glucose (LG, 5.5 mM/L) and high glucose (HG, 30 mM/L) for 48 h.