Anti cd73 fitc

Cell phenotype detection was performed as previously reported [38 (link)]. After reaching 80–90% confluence, P3 ADSCs were digested with 0.25% pancreatin-0.02% ethylenediamine tetraacetic acid (EDTA; T1300, Solarbio), and then no less than 2 × 10⁵ cells were collected from each sample for analysis. Next, cells were resuspended in 100 μL of PBS and subjected to centrifugation at 1,000 r/min for 5 min. After removing the PBS, cells were resuspended in 200 μL of PBS, incubated for 30 min with antibodies, and washed twice with PBS. After removing the PBS again and resuspending the cells with 300 μL PBS, the cell suspension was analyzed by flow cytometry (Accuri™ C6, BD Biosciences, USA). The following monoclonal antibodies used in this experiment were purchased from eBioscience™ of Thermo Fisher Scientific: anti-CD29-FITC (#11-0291-80), anti-CD90-FITC (#11-0909-42), anti-CD73-FITC (#11-0739-41), anti-CD105-FITC (#MA1-19594), anti-CD34-FITC (#11-0349-41), and anti-CD45-FITC (#11-9459-41) antibodies.

Five cell surface markers (Sca-1, CD90, CD73, CD34 and CD45) were examined. BMMSCs were trypsinized and counted to ensure each sample had more than 1×10⁵ cells. The cells were incubated with antibody anti-Sca-1(FITC) (eBioscience, USA), anti-CD90 (PE) (Biolegend), anti-CD73(FITC) (eBioscience, USA), anti-CD34(PE) (Biolegend, USA) and anti-CD45(PE) (BD Biosciences, USA) for 30 min at room temperature. Then cells were washed twice and suspended for flow cytometry.