Cck 8 detection reagent

eESCs were cocultured with SCM-198 (0, 50, 100, 200, 400 µM) or CD4⁺CD25⁺ Tregs, which were washed with PBS for three times after the pretreatment with E2 (100 nM), E2+SCM-198 (100 nM, 200 µM), PPT (5 nM), or PPT+SCM-198 (5 nM, 200 µM) for 48 h.
According to the instructions, add 10 µl CCK8 detection reagent (Dojindo, Japan) and 90 µl fresh culture medium to each well. Measure the absorbance at 450 nm wavelength to detect cell viability.

The cytotoxicity of EGCG loaded starch hydrogel/contact lens composites was evaluated based on the effects of contact lenses extracts on the proliferation of human corneal epithelial cells (HCECs, Seoul, Korea) through cell counting kit-8 (CCK-8) assay. 2.5 and 5 mg of different type contact lenses were respectively dipped in 10 ml DMEM/F12 (Sigma-Aldrich) containing 10% (v/v) fetal bovine serum (Sigma-Aldrich) and 1% (v/v) antibiotic/antimycotics (Sigma-Aldrich) in 37°C, 5% CO₂ incubator for 24 h to obtain contact lenses extracts with concentrations of 1:4 and 1:2 (Li et al., 2020 (link)). While the HCECs were plated into a 96-well plate at 2000/well and cultured in standard culture medium (DMEM/F12, 10% FBS, 1% antibiotic/antimycotics) for 24 h in 37°C, 5% CO₂ incubator. Then, the culture medium of all the wells was taken out and different group cells were cultured with the prepared contact lens extracts along with fresh standard culture medium, respectively. After culturing in 37°C, 5% CO₂ incubator for 1, 3, 5 and 7 days, CCK-8 detection reagent (Dojindo Laboratories Kumamoto, Japan) was added to measure the absorbance of the cells at 450 nm wavelength with SpectraMax M2 (Molecular Devices, United States).