Phycoerythrin pe conjugated mouse anti human cd73

DFCs, iDFCs and dDFCs were digested and then resuspended in PBS containing 10%FBS to a final concentration of 1*10⁷ cells/ml. Distribute 100 μl aliquots of the cell suspension into each tube. Add antibodies to cells and incubate for 20 min in the dark. Antibodies used were FITC-conjugated mouse anti- human CD34 (BD Biosciences, 555821; 20 μl), phycoerythrin(PE)-conjugated mouse anti-human CD73 (BD Biosciences, 550257; 20 μl), FITC-conjugated mouse anti-human CD105 (BD Biosciences, 561443; 5 μl) and FITC-conjugated anti-human Strol-1. Finally, flow cytometry(BD Biosciences) was used to analyze the stained cells.

For flow cytometry analysis, MSCs or Pio-MSCs were trypsinized and washed twice prior to resuspension in PBS containing 2% FBS and 1 mM EDTA. Cells were adjusted to 1 × 10⁶ in 100 µL of cell suspension. For cell surface labeling, cell suspensions were incubated at 4 °C for 30 min with 5 µL of antibodies (dilution, 1:20) against MSC-specific surface markers. Phycoerythrin (PE)-conjugated mouse anti-human CD73, fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD90, and PE-conjugated mouse anti-human CD105 antibodies were supplied by BD Biosciences (San Jose, CA, USA). FITC-conjugated mouse anti-human CD34 was supplied by BD PharMingenTM (San Jose, CA, USA). Cell surface marker analysis was performed using a BD FACSCanto™ II Flow Cytometer and FACSDIVA software version 6.1.3 (BD Biosciences, San Jose, CA, USA).

For flow cytometry analysis, MSCs or iMSCs were trypsinized and washed twice prior to resuspension in PBS containing 2% FBS and 1 mM EDTA. Cells were adjusted to 1 × 10⁶ in 100 µL of cell suspension. For cell surface labeling, cell suspensions were incubated at 4 °C for 30 min with 5 µL of antibodies (dilution, 1:20) against MSC and iPSC-specific surface markers. Phycoerythrin (PE)-conjugated mouse anti-human CD73, fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD90, and PE-conjugated mouse anti-human CD105 antibodies were supplied by BD Biosciences (San Jose, CA, USA). FITC-conjugated mouse anti-human CD34 and PE-conjugated mouse anti-human SSEA4 antibodies were supplied by BD PharMingen™ (San Jose, CA, USA). Cell surface marker analysis was performed using a BD FACSCanto™ II Flow Cytometer and FACSDIVA software version 6.1.3 (BD Biosciences, San Jose, CA, USA).