Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Invitrogen), as described in our previous study [38 (link)]. Briefly, cells were seeded in KBM-2 basal media in a 96-well plate overnight in a 37 °C incubator with 5% CO2. The cells were incubated with varying concentrations of PVP-I (0%, 0.01%, 0.05%, 0.1%, 0.5%, 1% and 2%) for 1 min, followed by three washes with 1X
PBS and supplemented with fresh KBM-2 medium. The cells were incubated for 24 h at 37 °C, followed by the addition of 100 μL of MTT reagent (5 mg/mL in PBS) to each well for 4 h at 37 °C. The supernatant was then aspirated, followed by the addition of 100 μL of cell lysis buffer (20% SDS in 50% DMF) for an hour. The absorbance was measured using a microplate reader (Synergy multi-mode reader, BioTek, Winooski, VT, USA) and the cell viability was expressed as a percentage over control and calculated using the formula (mean OD of treated cells/mean OD of untreated control cells) × 100 and expressed as cell viability (%).
PBS and supplemented with fresh KBM-2 medium. The cells were incubated for 24 h at 37 °C, followed by the addition of 100 μL of MTT reagent (5 mg/mL in PBS) to each well for 4 h at 37 °C. The supernatant was then aspirated, followed by the addition of 100 μL of cell lysis buffer (20% SDS in 50% DMF) for an hour. The absorbance was measured using a microplate reader (Synergy multi-mode reader, BioTek, Winooski, VT, USA) and the cell viability was expressed as a percentage over control and calculated using the formula (mean OD of treated cells/mean OD of untreated control cells) × 100 and expressed as cell viability (%).
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