Cellcounting lite 2.0 luminescent cell viability assay

The human NB cell lines (SK-N-SH, SH-SY5Y, SK-N-BE (2), and IMR-32) were obtained from the National Collection of Authenticated Cell Cultures in Shanghai, China.
SK-N-SH and IMR-32 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin–streptomycin mixture (Sigma-Aldrich). SK-N-BE (2) cells were cultured in DMEM/F12 (1:1) with 10% FBS and 1% penicillin–streptomycin mixture (Sigma-Aldrich). SH-SY5Y cells were cultured in a mixture of MEM (44.5%) and Ham’s F12 (44.5%), supplemented with 10% FBS (Gibco), 1% non-essential amino acids (NEAAs) (Gibco), and 1% penicillin–streptomycin mixture (Sigma-Aldrich). All cells were maintained in an incubator at 37°C with a humidified atmosphere containing 5% CO₂.
Gomisin B and ginsenoside Rh2 chemical reagents were procured from MedChemExpress. Gomisin B (purity ≥99.9%) and ginsenoside Rh2 (purity ≥99.9%) were dissolved in dimethyl sulfoxide (DMSO). To ensure minimal impact on cells, the final concentration of DMSO in the cell culture medium was maintained below 0.1%.
Cell viability was assessed using the CellCounting-Lite 2.0 Luminescent Cell Viability Assay (Vazyme, DD1101-03) and measured using a multimode microplate detection system (PerkinElmer EnVision 2015). Aidi injection was donated by Guizhou Ebay Pharmaceutical Co., Ltd.

Cells of about OD₆₂₀ = 0.5 were collected at intervals. Before cell viability detection, lyase was added to pretreat the cell for 2 h. Cell viability in the presence of acetic acid and formic acid in SD medium was assayed, followed by CellCounting-Lite 2.0 Luminescent Cell Viability Assay (Vazyme Biotech Co., Ltd., Nanjing, China). CellCounting-Lite2.0 used in this study is a cell viability detection reagent based on the luciferase system. The reagent contains high-purity luciferin and thermostable luciferase. When we added this product to the cell culture to lyse the cells and release ATP, and the reaction shown in the figure below can be produced, and a stable “light” type signal can be issued. The luminescence intensity is proportional to the amount of ATP, that is, the number of living cells within a certain range. So, we used cell viability to present relative ATP levels. Luminescence was measured by SpectraMax M2e (Molecular Device) at 560 nm. The data were expressed as mean ± SD. The bars in the figures indicate the ranges of the standard deviation.