Victor nivo multimode plate reader

Human colonic adenocarcinoma HCT116 and Caco-2 cells were cultured in the advanced DMEM containing 10 % (v/v) of FBS, and 1 % penicillin–streptomycin solution, and maintained at 37 °C under 5 % CO₂ humidified atmosphere. Cell viability was measured using MTT assay method. HCT116 and Caco2 cells were inoculated in 96-well plates at a density of 2 × 10⁴ per well. After incubated for 12 h, the cells were treated with free Fx (dissolved in DMSO with a final working concentration of <0.1 %), 2-HP-β-CD or Fx/2-HP-β-CD IC (dissolved in PBS) at corresponding concentrations. After 24 h incubation, the medium was replaced with 100 μL fresh DMEM containing 10 % MTT solution and incubated for 4 h. The medium was then removed and 110 μL of DMSO was added into each well to dissolve the formazan crystals. The absorbance at 570 nm was measured using a VICTOR Nivo multimode plate reader (PerkinElmer, USA). DMEM medium was used as the negative control group. The cell viability (CV) was calculated according to the following formula: $\mathit{CV}(\%)=\frac{A_1-A_0}{A^{\prime }-A_0}\times 100$ where A₁, A' and A₀ are the absorption values of experimental well, negative control well and blank well.