Cd11b alexa fluor 488

Characteristic surface markers (cluster of differentiation, CD) of the S-ASCs and V-ASCs from three animals were determined using flow cytometry (FACSCalibur; BD Biosciences, Mississauga, ON, Canada). Briefly, 10 6 live cells at the third to fifth passages were suspended in 100 µl wash flow buffer (WFB). The WFB contains 1× PBS, 2% FBS, 0.05% sodium azide (Sigma-Aldrich). Then the ASCs were incubated with antibodies of interest at 4°C for 30 min. The antibodies used in this study included CD11b-Alexa Fluor 488 (1:200 dilution, 201812 ; Biolegend, San Diego, CA, USA), CD29-Alexa Fluor 488 (1:50 dilution, 102212; Biolegend), CD31-Alexa Fluor 488 (1:10 dilution, MCA1334A488; AbD Serotec, Raleigh, NC, USA), CD45-phycoerythrin (PE) (1:100 dilution, 202207; Biolegend), CD59-fluorescein isothiocyanate (FITC) (1:20 dilution, 550976; BD Biosciences), CD90-FITC (1:200 dilution, 202504; Biolegend) , and CD106-PE (1:20 dilution, 200403; Biolegend) . For assessment of CD73 expression, the ASCs were incubated with primary antibody CD73 (1:50 dilution, 551123; BD Biosciences) at 4°C for 30 min and then washed with PBS three times and stained with second antibody goat anti-mouse IgG-FITC (1:100 dilution, sc-2010; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for 1 h. FACS was performed using 20,000 cells per sample.