Cck 8 assay kit

CSCs of OVCAR-3 and OVCAR-3/DDP (CSCs/DDP) were treated with 0, 10, 20, 40, and 80 µM of lumiflavin and 0, 40, 80, 160, 320 and 640 µM of DDP for 72 h. Surviving CSCs/DDP were compared with the CSCs combining cell viability/proliferation assays using the CCK-8 assay kit (Dojindo Laboratories, Japan) according to the manufacturer’s instruction. The sensitivity of these paired cell lines is usually determined by exposing them to a range of drug concentrations and then assessing cell viability. The IC₅₀ (drug concentration causing 50% growth inhibition) for these paired cell lines can be used to determine the increase in resistance known as the resistance index by the following equation (21 (link)):

Human podocytes were seeded at a density 3×10⁴ cells/cm² in RPMI 1640 medium on 96-well plates. After cell adherence, RPMI-1640 medium was replaced with other media, including NG, MG, HG, or HG + FA at concentrations of 2.5, 5, or 10 μg/mL. Cells were subsequently incubated for 48 h. Cell viability was detected using the CCK-8 assay kit (cat no. CK04-11; Dojindo Molecular Technologies, Inc., Kumamoto, Japan) according to the manufacturer’s instructions. The absorbance of each sample was measured at a wavelength of 450 nm.

The CCK‐8 assay kit (Dojindo) was used for testing cytotoxicity in vitro. At PD45, 2BS or WI38 cells were seeded into flat‐bottomed 96‐well microplates at a density of 5 × 10³ cells/0.2 ml per well. After 20 hr, when the cells reached a subconfluent state, the cells were transferred to a special culture medium containing various concentrations of BBR for further growth, at 37°C in 5% CO₂ up to 24 hr. Then, the CCK‐8 solution (diluted 0.1 times with 10% FBS DMEM) was added to each well and the cells were incubated for 1 hr at 37°C. The absorbance values of each well were determined spectrophotometrically at 490 nm using a microplate reader (Biotek, MQX200).
Cell viability (%) = (OD _{treatment group} − OD _blank)/ (OD_{control group} − OD _blank) × 100.
Cell proliferation was assayed using the CCK‐8 method. Cells were seeded into 96‐well plates at 2.5 × 10³ cells/0.2 ml per well and incubated with different concentrations of BBR (0, 0.3125, 1.25, and 5 μg/ml) for one week. The absorbance values of each well were measured at day 0 (4 hr after plating) and on days 1, 2, 3, 4, 5, and 6. The medium containing BBR or DMSO was refreshed every 24 hr. At the indicated points, cells were harvested with 10% CCK‐8 for one hour, and the absorbance was measured at 490 nm. Each data point was measured five times, and each curve was repeated more than three times.

Cell growth was detected with a CCK-8 assay kit (Dojindo, Japan) according to the manufacturer’s instructions. These cells were plated in 96-well plates and cultured for 0, 24, 48 and 72 hours. Ten microliters of CCK-8 were added to each well, and the cells were cultured for an additional 3 hours. The absorbance was detected at 450 nm at different time points. To analyze cell cycle progression, GC cells were stained with cell cycle reagent (Thermo) following a standard protocol. The cell cycle was measured with flow cytometry on a Beckman flow cytometer (Dickinson, USA). The protein levels of IL-1β, IL-6 and TNF-α in the cell suspension were detected by ELISA following the manufacturer’s protocol.