Cell counting kit 8 cck 8

The proliferation capacity of the trophoblast cells was assessed by Cell Counting Kit-8 (CCK-8) (Vazyme Biotech Co., Ltd., Nanjing, China) assay according to the manufacturer's instructions. The HTR-8/SVneo cells were plated into the 96-well plate after being transfected with lentiviruses overexpressing RPAIN or a negative control. These cells were cultured for 6 h with five replicate wells at 2,000 cells per well. The cells were cultured at 37°C with 5% CO₂. Then, 10 μl of the CCK-8 solution were added to the medium after the cells were incubated for 0, 24, 48, and 72 h. Finally, the absorbance was measured at 450 nm with a multifunctional microplate reader. Each experiment was performed in triplicate.

Cell proliferation was measured by using the Cell Counting Kit-8 (CCK-8) (Vazyme; A311-01) according to the manufacturer’s instructions. Briefly, cells were seeded onto plastic 96-well plates at an initial density of 2 × 10³ cells/well. Then, CCK8 solution was added to each well at the indicated times and incubated for an additional 2 h at 37°C. Thereafter, OD₄₅₀ values were measured.

Cell number was counted by the automated cell counter (Invitrogen, USA). Cell viability was determined using the Cell Counting Kit-8 (CCK-8) (Vazyme, Jiangsu, China) assay.

A density of 5000 cells/well was used to seed cells into 96-well plates, after which they were allowed to grow for 24 h. Next, 10 μl of Cell Counting Kit-8 (CCK-8) (Vazyme, Nanjng, China) was introduced to each well, followed by a 2-h incubation at 37 °C. The Eon™ Microplate Reader (BioTek, VT, USA) was used to determine the absorbance of each well at a wavelength of 450 nm.

The Cell Counting Kit-8 (CCK8) (Vazyme, Nanjing, China) was used to measure cell proliferation after the cells were harvested and dispensed into 96-well plate (at 1000 cells per well). The cells were seeded into a 6-well plate and incubated for 12 days at 5% CO₂, 37 °C, fixed in 4% paraformaldehyde, and stained with 0.1% crystal violet.