4.3. Cell Viability Assay

Cell viability was determined by MTT assay. B16F10 cells were plated in 96 well plates at a density of 3 × 10³ cells per well. When cells grow to 30% density, the cells were incubated with the compounds for 48 h, the culture medium was removed and replaced with 20 mL of MTT solution (5 mg·mL⁻¹) dissolved in fresh DMEM and incubated for 4 h. The medium was removed completely, and 150 µL of DMSO was added to each well and fully dissolved for 5 min. Optical absorbance was set at 570 nm with a microplate spectrophotometer (, New Jersey, USA). Absorbance of cells without treatment was considered as 100% of cell survival. Each treatment was performed in six multiple holes.