MTT assay for cell viability

The MTT assay is a standard method used to assess cell viability. Briefly, H9c2 cells were washed twice with PBS, detached with 0.25% trypsin and made into a single cell suspension. Cells were seeded into 96-well microtiter plates at a density of 3-5×10³ cells/well in 200 µl culture medium. Following incubation with 4 mM NAC for 1 h at 37°C, the cells were treated with 0.75 mM H₂O₂ and were incubated for various durations. Subsequently, 100 µl MTT solution (0.5 mg/ml) was added to each well and the cells were incubated for 4 h at 37°C, after which, 150 µl dimethyl sulfoxide was added to each well. The absorbance was measured at 570 nm, and the values were used to calculate the relative ratio of viable cells to total cells.