Cell Viability Assay Protocol

The assay was adapted from a previous report [21 (link)]. Briefly, treated cells (5,000 cells/well) were inoculated into 96 wells culture plate in triplicate. The cells were maintained at 37°C for the indicated times. After 0, 24, 48, and 72 h, CCK-8 reagent (Sigma) was added to the cells to cultivate cells for two more hours. Finally, the absorbance of each well was measured at a wavelength of 450 nm using a microplate reader (Thermo Fisher Scientific). A cell viability curve was generated by plotting the optical density (OD₄₅₀) values against time.

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Xu H, & Li B. (2022). MicroRNA-582-3p targeting ribonucleotide reductase regulatory subunit M2 inhibits the tumorigenesis of hepatocellular carcinoma by regulating the Wnt/β-catenin signaling pathway. Bioengineered, 13(5), 12876-12887.

Publication 2022

CCK-8 reagent microplate reader

Assay Cck 8 Cell viability Cells Optical

Corresponding Organization : Huangshi Central Hospital

Other organizations : Wuhan General Hospital of Guangzhou

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Variable analysis

independent variables

Treatment condition(s)

dependent variables

Cell viability

control variables

Cell seeding density (5,000 cells/well)
Incubation temperature (37°C)
Incubation duration (0, 24, 48, and 72 h)
CCK-8 reagent addition and incubation time (2 h)
Absorbance measurement wavelength (450 nm)

controls

The assay was adapted from a previous report [21 (link)].

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