Cell Viability Assays Comparison

Viability of cells was determined using the Cell Titer 96® AQueous One Solution Cell Proliferation Assay (Promega, Mannheim, Germany) as reported [56 (link)]. Cells were grown overnight in 96-well plates and then treated with increasing concentrations of OH-ME as indicated. DMSO served as solvent control. After 72 h, viability was measured using a microplate reader (Sunrise Tecan Reader, Crailsheim, Germany) according to the manufacturer’s instructions. Viability was also assessed with the CellTiter-Glo®Luminescent Cell Viability Assay (Promega, Mannheim, Germany), which measures the cellular ATP level. To this end, the cells were seeded in white 96-well-plates, treated as indicated, and incubated for 72 h. The assay was performed according to the manufacturer’s instructions in the multiwell reader Fluoroskan Ascent FL (Thermo Scientific). In addition, an AlamarBlue assay was performed. Cells were seeded in 96-well plates, treated as indicated, and incubated for 72 h. Incubation medium was exchanged with DMEM without supplements, containing 44 µM AlamarBlue and cells were incubated for 90 min. Cell viability was measured fluorometrically with excitation wavelength λ = 544 nm and emission wavelength λ = 590 nm (Fluoroskan Ascent FL, Thermo Scientific).