Pe conjugated mouse anti human cd73

hPDLSCs and hGMSCs were characterized by analyzing the expression of characteristic cell surface markers. Single cell suspensions were stained 1 : 10 with one of the following antibodies (all from eBioscience, San Diego, USA) for 20 min: phycoerythrin- (PE-) conjugated mouse anti-human CD90, PE-conjugated mouse anti-human CD105, PE-conjugated mouse anti-human CD146, PE-conjugated mouse anti-human CD29, PE-conjugated mouse anti-human CD73, fluorescein isothiocyanate- (FITC-) conjugated mouse anti-human CD34, FITC-conjugated mouse anti-human CD45, and FITC-conjugated mouse anti-human CD31. After resuspending cells in 200 μl of FACS buffer (3% BSA, 0.09% sodium azide, in 1x PBS), an argon laser was used, exciting the fluorescence at 488 nm. The percentage of positive cells for each investigated surface marker was determined using the FACScan Flow Cytometer (Becton Dickinson, Franklin Lakes, USA).

Phycoerythrin (PE)-conjugated mouse anti-human CD73, fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD90, PE-conjugated mouse anti-human CD105, and FITC-conjugated mouse anti-human CD34 antibodies were purchased from eBioscience (Waltham, MA, USA). MACS Plex Exosome Kit (Miltenyi Biotec, Bergisch Gladbach, Germany) was used to evaluate the exosome surface markers (CD63 and CD81). PE-conjugated mouse anti-human IDO was treated after permeabilization with 0.1% Triton X-100 in PBS for 10 min. All data were measured with the Acoustic Focusing Cytometer (Thermo Fisher Scientific, Waltham, MA, USA).