Model elx800 microplate reader

Cell viability and proliferation were measured according to the protocol of the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) Cell Proliferation and Cytotoxicity Assay Kit (Beyotime Biotechnology, China). UC-MSCs at passage 4 were seeded in 96-well plates and incubated for 2, 3, 4, and 5 days. MTT was added to each well, and UC-MSCs were incubated at 37 °C in the dark for 4 h. MTT was then removed, and 150 μL of DMSO was added to each well. Absorbance at 570 nm was measured using a Model ELX800 microplate reader (Bio-Tek Instruments). All experiments were performed in triplicate.

The cell metabolism rate of the cell lines was measured using the MTT assay. Exponentially growing cells were treated for 24 h or 48 h with various concentrations (0–500 μmol/L) of bicyclol in 96-well plates. DMSO-treated cells (0.25 %) were used as vehicle controls. MTT was then added to each well, and the cells were incubated for 4 h at 37 °C in the dark. The Formazan crystals that formed were dissolved with 150 μl of DMSO. The absorbance at 570 nm was measured using a Model ELX800 microplate reader (Bio-Tek Instruments). Each test was repeated at least three times. The cell metabolism rate was calculated by the following formula: %cell metabolism rate = (mean absorbance in test wells)/(mean absorbance in control well) x 100 %.