Cell Viability Assay with MTT

To assess cell viability, we used the cell proliferation kit I (MTT) (Sigma-Aldrich, Castle Hill, NSW, Australia) following the procedures described in previous work [50 (link)]. Cells were seeded into 96-well plates at a concentration of 1 × 10⁴ cells/well. DMEM containing 0.5 mg/mL 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) solution was added in each well. Following incubation for 6 and 24 h at 37 °C, the medium was removed, and 100 μL of solubilization solution was added. Formazan formed by the cleavage of the yellow tetrazolium salt MTT was measured spectrophotometrically by absorbance change at 550–600 nm using a microplate reader.