Quantifying Cell Metabolic Activity in Scaffolds

The metabolic activity of cells in scaffolds was quantitatively evaluated by mitochondrial dehydrogenase activity using MTS assay (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) from BioVision (Catalog #K300-500), according to the manufacturer’s protocol. Scaffolds were located in 48-well plates. Cells were seeded onto the scaffolds (0.75 × 10⁴ cells in 400 mL per well) as per the ratio of 10% MTS assay into cell-scaffold media and incubated for 4 h. The cell scaffolds were cultured from days 3, 5, 10, and 30 at 37 °C/5% CO₂. After incubation for an hour, the cultures were measured at 490 nm [27 (link)].