Cyquant mtt cell viability assay kit

MTT assay was used for cell viability analysis by CyQUANT™ MTT Cell Viability Assay Kit (Invitrogen, V13154). 1 × 10⁴ SiHa and HeLa cells were respectively seeded into the 96-well plates overnight. 10 μL MTT solution was incubated to cells after transfection for 0 h, 24 h, 48 h and 72 h. 4 h later, 100 μL dimethyl sulfoxide (DMSO; Invitrogen) was added to dissolve the formazan. The optical density (OD) value of 490 nm was read under the microplate reader (Thermo Fisher Scientific).

Fibroblasts were seeded in six-well plates (3 × 10⁵ cells/well) with a base medium. The fibroblasts were treated with PM₁₀ of various PM concentrations (1, 5, 10, 50, and 100 µg/mL) and incubated for 72 h. To assess cell proliferation, the cells were incubated with a 0.05% trypsin solution for 3 min. Cells were suspended using base medium, and then mix 10 µL of cell with 10 µL of trypan blue, and pipet into a disposable countess chamber slide. Total cells were counted using the Countess™ 3 Cell Counter (Invitrogen, Waltham, MA, USA). As a result, To evaluate the cell viability, fibroblasts were seeded in triplicate in 96-well plates (5 × 10³ cells/well) with a base medium. The fibroblasts were treated with PM₁₀ of various PM concentrations (1, 5, 10, 50, and 100 μg/mL) and incubated for 24, 48, and 72 h. To determine cell viability, an MTT assay was performed using CyQUANT™ MTT Cell Viability Assay Kit (Invitrogen, Waltham, MA, USA) according to the manufacturer’s protocol. Absorbance was measured at 570 nm using a microplate spectrophotometer (Epoch 2; BioTek, Winooski, VT, USA).

Cell viability assay was performed with CyQUANT MTT Cell Viability Assay Kit (Invitrogen-Life Technologies Inc., Gaithersburg, MD, USA). The cells were seeded in 96-well plates (1 × 10⁴ cells per well) and incubated at 37 °C. Different concentrations of AST were added to the cells exposed to high glucose. After 24 h incubation, 5 mg/mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was added to each well and incubated at 37 °C for 4 h. The culture medium supernatant was removed, and the formazan was dissolved with dimethyl sulfoxide (DMSO) for 30 min at 25 °C. The absorbance was measured at 570 nm with a microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA).