The Cell Counting Kit‐8 (CCK‐8) reagent (Nanjing Jiancheng Bioengineering) assay has the advantages of excellent repeatability and low cytotoxicity. HaCaT cells at the logarithmic (log) phase of 4–5 generations were harvested and inoculated into 96‐well plates using 100 μL of (4–5) × 104 cells/mL.
14 After the cells were cultured at 37°C for an overnight period in 5% CO2, the 96‐well plates were incubated with varying concentrations of LMWP. After 24 h of interaction between LMWP and the cells, CCK‐8 reagent was added at a concentration of 10% (V/V), and the cells were incubated in the cell incubator (bluepard) for approximately 2 h. Finally, the OD value at 450 nm was determined using a microplate reader (MultiSKAN), and each concentration was tested in five replicates. The cell proliferation rate is calculated strictly based on the following formula
15 (link):
14 After the cells were cultured at 37°C for an overnight period in 5% CO2, the 96‐well plates were incubated with varying concentrations of LMWP. After 24 h of interaction between LMWP and the cells, CCK‐8 reagent was added at a concentration of 10% (V/V), and the cells were incubated in the cell incubator (bluepard) for approximately 2 h. Finally, the OD value at 450 nm was determined using a microplate reader (MultiSKAN), and each concentration was tested in five replicates. The cell proliferation rate is calculated strictly based on the following formula
15 (link):