Pe anti mouse cd90

After culture for 14 days, the differentiated rBMSCs were harvested using 0.25% trypsin/EDTA. A 100-μL cell suspension with a cell density of 1 × 10⁵/mL was transferred into an Eppendorf tube and then incubated with the following antibodies: a primary antibody CD31 (ab9498, Abcam, dilution 1:500) and PE anti-mouse CD90 (BD, USA) for 30 min at 4°C. A nondifferentiated rBMSC group served as a control. Then the stained samples were assessed by a flow cytometer (FACSVerse, BD, USA) and analyzed by FlowJo software. We designated the CD31-positive expression cells as induced endothelial cells (iECs). Platelet endothelial cell adhesion molecule (PECAM-1), also known as CD31, is a protein that is normally found on endothelial cells. CD90 can be used as a marker for a variety of stem cells.

Flow cytometry was performed for analysis of cell surface antigen expression. The MSCs at passage one were detached using Accutase (Innovative Technologies Inc., San Diego, CA, USA) and resuspend into PBS at a concentration of 1 × 10⁵ cells per 100 µL. The cells were then incubated with fluorescence-labeled primary antibody or isotype control for 30 min in dark condition. The utilized primary antibodies were: FITC anti-mouse Ly-6A/E Sca-1 (Biolegend), PE anti-mouse CD 90, PE anti-mouse CD44, PE anti-mouse CD 146, FITC anti-mouse CD14, PE anti-mouse CD34 (BD, Franklin Lakes, NJ, USA). These samples were analyzed by Accuri™ C 6 Flow Cytometer (BD).