MTT Assay for PC12 Cell Viability

Pheochromocytoma PC12 cells, derived from rat adrenal medulla, were obtained from American Type Culture Collection (Manassas, VA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 6% fetal bovine serum and horse serum, 100 units/mL penicillin and 100 μg/mL streptomycin in a humidified CO₂ (7.5%) incubator at 37°C. Fresh medium was applied every other day. Culture reagents were from Invitrogen (Carlsbad, CA). Cell viability was assessed by MTT [3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide] assay. Cells were plated in 96-well plate for 24 hours and treated with drugs for 48 hours before adding MTT. Then the cells were incubated with MTT for another 3 hours at 37°C. After that, absorbance of 570 nm was measured in a microplate reader (Thermo Fisher Scientific, Waltham, MA) [10 (link)].