Cell Viability Assay for hWJ-MSCs

hWJ-MSCs seeded in 6-well plates were exposed for 24 h to CB (range 0.01–5 μM) or DMSO. Each experimental condition was assayed in technical duplicate. At the end of the exposure-time, CTR and treated cells were detached by trypsin-EDTA and counted, as previously described [22 (link)]. Briefly, cells were resuspended in a medium with 50% of Erythrosine B red dye 0.2% (Sigma-Aldrich Co., St. Louis, MO, USA) in PBS. Not stained viable cells and red stained dead cells were manually counted, at least twice for each condition, using the Neubauer hemocytometer (BRAND GmbH, Wertheim, Germany) and a light microscope (Leica Labovert FS Inverted Microscope, Wetzlar, Germany). The total number of viable and dead cells was calculated according to the manufactured instructions.