Cell viability was determined by MTT test as described earlier [54 (link)]. Cells were seeded in a 96-well plate in the amount of 1 × 104 cells/200 µL and cultured at 37 °C in a humidified CO2 atmosphere (5%) in a nutrient medium DMEM (PanEco, Moscow, Russia) and MEM (PanEco, Moscow, Russia). After 24 h of incubation, 2 mL aliquots of test compounds (0.1 to 100 mM) dissolved in DMSO were added to the cell cultures, and cells were cultured under the same conditions for 24 h. The final content of DMSO in the well did not exceed 1% and did not have a toxic effect on the cells. DMSO also was added to the control wells in a volume of 1%.
After 24 h, 20 mL of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5 mg/mL) was added to each well and the plates were additionally incubated for 2 h.
Using a plate analyzer (Cytation3, BioTek Instruments Inc., Winooski, VT, USA), the optical density was determined at 530 nm. The concentration value causing 50% inhibition of cell population growth (IC50) was determined from dose-dependent curves.
After 24 h, 20 mL of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5 mg/mL) was added to each well and the plates were additionally incubated for 2 h.
Using a plate analyzer (Cytation3, BioTek Instruments Inc., Winooski, VT, USA), the optical density was determined at 530 nm. The concentration value causing 50% inhibition of cell population growth (IC50) was determined from dose-dependent curves.
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