Viability was assayed using CellTiter-Blue® cell viability assay kit and performed as per manufacturer’s instructions. For viability assay, MSCs in the co-culture were inactivated by 10 μg/mL mitomycin C (Dalian Meilun Biology Technology Co., Ltd, China) for 3 h before being mixed with mFBs. Cells were plated in 48-well plate with constant number of mFBs and an equal number of inactivated MSCs used in the co-culture among each ratio were plated in individual wells for background elimination. Before the assay, culture medium was discarded. Alamar blue solution prepared in fresh medium was added to each well and incubated at 37°C. Fluorescence values were read by microplate reader (Molecular Devices, USA) with excitation at 560 nm and emission at 590 nm. All values were normalized to the mFB mono cultures.
Cell apoptosis detection kit (Beyotime, China) was used to verify that the mFBs were viable in co-culture. All procedures were accomplished as per the instructions. Cells treated with 500 μmol/L H2O2 for 2 h were used as positive control.
Cell apoptosis detection kit (Beyotime, China) was used to verify that the mFBs were viable in co-culture. All procedures were accomplished as per the instructions. Cells treated with 500 μmol/L H2O2 for 2 h were used as positive control.