This assay was performed to determine the median inhibition concentration on cancer cell of A375. Cell viability to determine 50% inhibitory concentration (IC50) and proliferation assay were evaluated using Promega CellTiter 96® AQueous Non-Radioactive Cell Proliferation (Promega, Madison, WI, US) assay as recommended by the manufacturer. In determining IC50, cells were seeded at 4.0 × 102 cells/mL and treated with PB extract with serial concentrations of 3.9–1000 µg/mL in a 96-well plate before incubation for 72 h. Then, 10 µL of MTS reagent were added in each well and incubated for 3 h before measuring at 490 nm using a microplate reader. The IC50 values for PB aqueous extract and 5-FU were calculated using Graphpad Prism 6. Further test for the proliferation assay over time (24, 48, 72 and 95 h) used untreated (UT) cells as negative control and 5-fluorouracil as positive control. Additionally, the IC50 of PB aqueous extract was evaluated on NHDF with UT and 5-FU as controls.
Celltiter 96 aqueous non radioactive cell proliferation
Manufactured by Promega
Sourced in United States
The CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The assay uses a tetrazolium compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron coupling reagent (phenazine methosulfate; PMS) to produce a formazan product that is soluble in tissue culture medium. The absorbance of the formazan product is measured at 490nm, which is directly proportional to the number of living cells in culture.
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3 protocols using celltiter 96 aqueous non radioactive cell proliferation
1
Anticancer Potential of PB Extract
2
Optimized Scaffold Design for hMSC Adhesion
3
Nanoparticle Cytotoxicity Evaluation in BJ5ta Cells
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Cell viability was studied using the Promega CellTiter 96 ® AQueous Non-Radioactive Cell Proliferation (MTS) assay. BJ5ta cells were seeded in 96-well tissue culture polystyrene plates at a density of 1 × 10 4 cells/well and incubated overnight to promote cell adhesion. The cells were incubated with different concentrations of nanoparticles (from 0.0625 mg/mL up to 1 mg/mL). The concentrations of nanoemulsions for the application in cells were calculated based in the concentration of cyclo-oligosaccharides in the initial formulations (5 mg/mL). The nanoemulsions were incubated with cells for 24, 48 and 72 h. A MTS mixture was then added and the cells were further incubated for 4 h at 37 • C. After this period, the plates were placed on Synergy Mx Multi-Mode Reader from BioTek (USA) and the absorbance of the formazan product was read at 490 nm. Cell viability was expressed as a percentage relative to the negative control (untreated control cells). Two independent experiments were made.
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