Percp cy 5.5 mouse anti human cd73

Normoxic hWJ-MSCs treated with rFGF-17 and transfected with siFGF-17 at passage 7 or hypoxic hWJ-MSCs treated with rFGF-17 and transfected with siFGF-17 at passage 10 were harvested and washed with 1×PBS (Intron Biotechnology, Seoul, Korea). Normoxic hWJ-MSCs not treated with rFGF-17 and transfected with negative control siRNA or hypoxic hWJ-MSCs not treated with rFGF-17 and transfected with negative control siRNA were used as respective control groups. Cells were fixed with BD Cytofix Fixation Buffer (BD Biosciences, Piscataway, NJ, USA) and stained with V450 mouse anti-human CD31 (1:20), fluorescein isothiocyanate (FITC) mouse anti-human CD34 (1:20), phycoerythrin (PE)-Cy^™7 mouse anti-human CD44 (1:20), V500 mouse anti-human CD45 (1:20), PerCP-Cy^™5.5 mouse anti-human CD73 (1:20), PE mouse anti-human CD90 (1:20), APC mouse anti-human CD105 (1 : 20), and V450 mouse anti-human CD144 (1:20) (BD Biosciences) antibodies for 30 min at room temperature. Cells were washed twice with Stain Buffer (BD Biosciences) and analyzed with a FACSVerse^™ flow cytometer (BD Biosciences) and Flowjo software (Treestar, San Carlos, CA, USA).

Normoxic and hypoxic hUC-MSCs at passage 6 were harvested, fixed with BD Cytofix Fixation Buffer (BD Biosciences, Piscataway, NJ, USA) for 10 min at room temperature and washed with Perm/Wash buffer (BD Biosciences). Cells were stained with V450 mouse anti-human CD31 (1:20), fluorescein isothiocyanate (FITC) mouse anti-human CD34 (1:20), phycoerythrin (PE)-Cy^™7 mouse anti-human CD44 (1:20), V500 mouse anti-human CD45 (1:20), PerCP-Cy^™5.5 mouse anti-human CD73 (1:20), PE mouse anti-human CD90 (1:20), APC mouse anti-human CD105 (1:20), or V450 mouse anti-human CD144 (1:20) (BD Biosciences) antibodies for 30 minutes at room temperature. Cells were washed with Stain Buffer (BD Biosciences), and analyzed with a Caliber flow cytometer (BD Biosciences) using CellQuest program (BD Biosciences) and Flowjo software (Treestar, San Carlos, CA, USA).