Cell Counting Kit-8

Protocol tips

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	- Cells were then incubated at 37°C for the next 7 days with ctrl, NG, HG, and HG + Met mediums. At day 1, day 3, and day 7 after seeding of the cells, CCK8 assay was performed

Protocol tips
- Cells were then incubated at 37°C for the next 7 days with ctrl, NG, HG, and HG + Met mediums. At day 1, day 3, and day 7 after seeding of the cells, CCK8 assay was performed

Publication protocol

Cell proliferation was analyzed using Cell Counting Kit-8 (CCK-8) assays. MG63 human osteoblast-like cells were plated in 96-well plates (1 × 103 cells/well). Cells were then incubated at 37°C for the next 7 days with ctrl, NG, HG, and HG + Met mediums. At day 1, day 3, and day 7 after seeding of the cells, CCK8 assay was performed according to the manufacturer's instructions. The optical density (OD) for each well was measured at 450 nm using a 96-well plate reader. Three replicate wells were used for each analysis and at least three independent experiments were performed. The cell proliferation rate was calculated according to the following equation provided by the manufacturer: the cell proliferation rate = [(OD experiment − OD blank)/(OD control − OD blank)] × 100% [11].

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