Immunophenotyping of Dental Pulp MSCs

In order to evaluate the expression of the typical mesenchymal stem cells (MSCs) markers, immune-selected hDPSCs, both in static and dynamic culture conditions, underwent FACS analysis against CD73, CD90, CD105, CD34, CD45, HLA-DR, as formerly described by Conserva et al. [19 (link)]. Following trypsin dissociation, cells were resuspended in culture medium and were stained with the following fluorochrome-conjugated antibodies (Abs): anti-human-CD73-PE-CY7, anti-human-CD90-FITC, anti-human-CD105-APC, anti-human-CD45-PE, and anti-human-HLADR-PE-CY7 (all from BD Biosciences, Franklin Lakes, NJ, USA); and anti-human-CD34-ECD (Beckman Coulter, Fullerton, CA, USA). A minimum of 10,000 cells per sample was acquired and analyzed by using the Attune Acoustic Focusing Flow Cytometer (Attune NxT, Thermo Fisher, Waltham, MA, USA). Data were analyzed by FlowJo 9.5.7 (Treestar, Inc., Ashland, OR, USA) under MacOS 10.