Inverted fluorescence microscopy system

Cell viability was determined using the Cell Counting Kit-8 (CCK-8) assay in 96-well plates containing 7000 cells per well. The DFCs were exposed to varying concentrations of H₂O₂ for 3, 6, and 12 h to determine the optimal H₂O₂ concentration for establishing a model of oxidative stress. Cell viability changes were observed using different concentrations of EVs co-cultured with cells for 24 h. To detect the protective effects of EVs on H₂O₂-induced DFCs, DFCs were pretreated with varying concentrations of EVs for 24 h and then exposed to 200 M H₂O₂ for 24 h to observe changes in cell activity. Then, 10% (v/v) CCK-8 medium was co-incubated with cells for 1 h to detect alterations in cell viability. Using a microplate scanner set at 450 nm, the optical density was calculated.
The ethynyldeoxyuridine (EdU, Ribo Bio, Guangzhou, China) assay was used to measure the percentage of DNA-replicating cells, which reflects the state of cell proliferation. With three independent samples for each treatment group, the ratio of the number of EdU-incorporated cells to the number of Hoechst-stained cells was used to determine the EdU incorporation rate. The specimens were visualised utilising an inverted fluorescence microscopy system (Olympus).

Cell viability was determined using the Cell Counting Kit-8 (CCK-8) assay in 96-well plates containing 7000 cells per well. The DFCs were exposed to varying concentrations of H₂O₂ for 3, 6, and 12 h to determine the optimal H₂O₂ concentration for establishing a model of oxidative stress. Cell viability changes were observed using different concentrations of EVs co-cultured with cells for 24 h. To detect the protective effects of EVs on H₂O₂-induced DFCs, DFCs were pretreated with varying concentrations of EVs for 24 h and then exposed to 200 M H₂O₂ for 24 h to observe changes in cell activity. Then, 10% (v/v) CCK-8 medium was co-incubated with cells for 1 h to detect alterations in cell viability. Using a microplate scanner set at 450 nm, the optical density was calculated.
The ethynyldeoxyuridine (EdU, Ribo Bio, Guangzhou, China) assay was used to measure the percentage of DNA-replicating cells, which reflects the state of cell proliferation. With three independent samples for each treatment group, the ratio of the number of EdU-incorporated cells to the number of Hoechst-stained cells was used to determine the EdU incorporation rate. The specimens were visualised utilising an inverted fluorescence microscopy system (Olympus).