The rat cardiomyocyte-derived H9c2 cell line (American Type Culture Collection) was cultured in high glucose-DMEM (GIBCO, Waltham, MA, USA) containing 10% FBS (Atlas Biologicals, Fort Collins, CO, USA) and 1% antibiotics (GIBCO). To induce necroptosis and apoptosis, cells were incubated in a hypoxic chamber (Thermo Fisher Scientific) with 1% O2, 5% CO2, and 94% N2. After exposure to hypoxia, cell counting kit solution (CCK-8, Dogen, Seoul, Korea) was added to each well at a final concentration of 0.5 mg/mL and incubated for 2 h at 37°C. Cell viability was determined by measuring cells at 450 nm.
H9c2 cell line
Manufactured by Thermo Fisher Scientific
Sourced in United States
The H9c2 cell line is a subclone of the original clonal cell line derived from embryonic rat heart tissue. It is a widely used in vitro model for the study of cardiomyocyte-like cells.
Automatically generated - may contain errors
2 protocols using h9c2 cell line
1
Hypoxia-Induced Necroptosis and Apoptosis
2
In vitro MIRI model in H9c2 cells
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The rat cardiomyocyte H9c2 cell line (American Type Culture Collection) was cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 1% streptomycin (100 µg/ml) and 1% penicillin (100 U/ml) at pH 7.4 in a 5% CO2 incubator at 37˚C. Hypoxia-reoxygenation (HR) was used to construct an in vitro model of MIRI in H9c2 cells. To mimic myocardial I/R in vitro, H9C2 cells at 80% confluence were incubated in DMEM without FBS (previously bubbled with gas mixture containing 95% N2 and 5% CO2) for 6 h at 37˚C. The cells were provided with fresh medium and then moved to 95% O2/5% CO2 for reoxygenation. The control plates were kept in the incubator with 95% O2/5% CO2 at 37˚C. Cells were harvested 16 h post-reoxygenation for analysis.
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