Cell Culture and Oxygen Glucose Deprivation/Re-Oxygenation (OGD/Re) Model

The rat cardiomyocyte-like H9c2 line was cultured in DMEM medium (11-995-040, Gibco) supplemented with 10% (v/v) fetal bovine serum (FBS) (SH30071.03, Hyclone), 1% penicillin/streptomycin (v/v), (15-140-122, Gibco) and 2 mM L-glutamine (25-030-081, Gibco). The cells were cultured in a humidied under 5% CO2 and 95% N2 at 37°C in a standard incubator and reached about 80% for experiments.

The H9c2 cardiomyocytes were pre-treated with of MS extract (6.0 µg/mL) for 24 h and then for OGD/Re (Zheng et al., 2017). Oxygen glucose deprivation condition was created by incubating the cells in glucose-free medium and an airtight Plexiglas chamber with an atmospheric composition of 5% CO₂ and 95% N₂ at 37°C for 3 h. After exposure to oxygen glucose deprivation for 3 h, the cells were reoxygenated by incubating in a standard 5% CO₂ incubator for 3 h.