Calcein am cell viability assay kit

Cell metabolism was studied after labeling hMSCs with SiC nanomaterials using CellTiter 96^® AQueous One Solution cell proliferation assay (MTS, Promega). After co-incubation with SiC nanomaterials for 4 hours, 20 μL of the assay reagent was pipetted into the samples in 100 μL of growth media. This was allowed to incubate under standard conditions for 4 hours. We then transferred 80 μL of the sample solutions to a new plate and read the absorbance at 490 nm on a spectrophotometer (SpectraMax M5, Molecular Devices). Cell viability was studied with the calcein AM cell viability assay kit (Biotium). Labeled cells were incubated with 100 μL of the reagent assay (containing 2 μM calcein AM) for approximately 1 hour and then the fluorescence was read at 517 nm with an excitation at 494 nm on a spectrophotometer (SpectraMax M5, Molecular Devices). Quantitative ROS measurement was performed by staining cells with 2’,7’-dichlorofluorescin diacetate (DCFDA, Sigma Aldrich) assay. H₂O₂-treated cells and nontreated cells were included as positive and negative controls, respectively. The fluorescence intensity was read at 530 nm with an excitation at 485 nm on a spectrophotometer (SpectraMax M5, Molecular Devices).

Cells were seeded at 2 × 10³/well in 96-well cell culture plates overnight, and incubated with varying concentrations of ADCs for 96 hours. Cell viability was determined using a Calcein-AM cell viability assay kit (Biotium Inc.) (10 (link)).