Phenotypic Characterization of Cell Cultures

Cultured cells were harvest, non specific binding was blocked and the cells were stained with different combinations of the following directly-conjugated antibodies: CD105 (cat no. 561443, BD Pharmingen), CD73 (cat no. 550257, BD Pharmingen), CD90 (cat no. 559869, BD Pharmingen), CD34 (cat no. 555821, BD Pharmingen), CD14 (cat no. 345785, BD), CD19 (cat no. 555415, BD Pharmingen), CD31 (cat no. 555445, BD Pharmingen), CD45 (cat no. 345808, BD), CD146 (cat no. 550315, BD Pharmingen), HLA-DR (cat no. 555558, BD Pharmingen) and HLA class I (cat no. 555553, BD Pharmingen) as described before5 (link). 7-amino-actinomycin D (Sigma Aldrich) was used for dead cell exclusion. Samples were analyzed on a FACSCalibur (BD Bioscience) and data acquisition and analysis were performed using CellQuest (BD Bioscience) and FlowJo software (Tree star, Ashland, Oregon, USA) respectively. Results are presented as percent positively stained cells.