Comprehensive MSC Surface Marker Profiling

The DPMSCs, UCMSCs, and ADMSCs were checked for their surface marker profile by FACSCalibur flow cytometry system (Becton Dickinson, CA, USA) as already described [8 (link), 9 (link), 23 (link)]. Briefly, the MSCs were detached from the surface of the flask with enzyme digestion for 3 minutes at room temperature, collected, and centrifuged at 300g for 5 minutes. The pellets were resuspended in stain buffer and the cells were counted by hemocytometer. Then, 2.5 × 10⁵ cells were incubated for 45 min, in the dark at 4°C, with the following antibodies: fluorescein isothiocyanate- (FITC-) labeled mouse antihuman CD90 (StemCell Technologies, Milan, Italy), CD105, CD14, and CD19 (Diaclone, France); R-phycoerythrin- (PE-) labeled mouse antihuman CD34, CD44, CD45 (Diaclone, France), and CD73 (Becton Dickinson, CA, USA); and anti-HLA-DR (Diaclone, France). The control for FITC- or PE-coupled antibodies was isotypic mouse IgG1. The data were evaluated using CellQuest software (Becton Dickinson, CA, USA).