Evaluating Cytotoxicity of Pharmaceutical Compounds

To determine the IC₅₀ of sunitinib (MedChemExpress; cat. no. HY-10255A), BAY-876 (MedChemExpress; cat. no. HY-100017) and NaSH on 786-O viability, cells were seeded in a 96-well culture plate (4x10³ cell/well) and incubated at 37˚C in a humidified incubator with 5% CO₂. After 24 h, the cells were treated with various concentrations of sunitinib (range, 1-100 µM), BAY-876 (range, 10-1,000 µM) and NaSH (range, 10-10³ µM) in quadruplicate for each concentration. The plates were incubated for 24, 48 and 72 h at 37˚C with 5% CO₂. For BAY-876 another plate was incubated for 96 h. After each incubation interval, the culture media of the plate wells were changed with new media and then the cell cytotoxicity was measured by using Cell Counting Kit-8 (CCK-8; MedChemExpress; cat. no. HY-K0301) (24 (link)) according to the manufacturer's instruction. Briefly, 10 µM of the CCK-8 reagent was added to each well, incubated for 2-4 h at 37˚C and then the absorbance was measured at 540 nm by using an ELIZA reader (BioTek Instruments, Inc.). Each experiment was carried out three times.