Cd31 pe cytm7

Fluorescence-activated cell sorting was used to analyze the cell surface markers in the SVF cell suspensions using the following monoclonal antibodies: mouse anti-human CD45-PE-Cytm5, CD31-PE-Cytm7, CD34-FITC, CD146-PE, CD13-PE, and CD73-FITC (BD Biosciences). The single-cell suspension was placed in a polystyrene tube on ice. An LSR II (BD Biosciences) flow cytometer was used to detect the cell density and cell surface markers in the SVF suspension.

For flow cytometry analyses, the cells were incubated with CD31-PE-Cy^TM7 (PECAM-1); CD45-PE-Cy^TM7 (leukocyte common antigen, Ly-5); CD11b-PE-Cy^TM7 (integrin αM chain); CD34-PE-Cy^TM7 (BD Biosciences, San Jose, CA); CD29-PE, Sca1-APC (Miltenyi Biotec, San Diego, CA); and CD105-PE (eBioscience, San Diego, CA) (1:200) to label hematopoietic, endothelial, and mesenchymal lineages. For cell viability, Sytox Blue 1/1000 (Invitrogen, Thermo Fischer Scientific, Waltham, MA) was added. Analyses were performed using BD LSR Fortessa SORP (BD Biosciences, San Jose, CA). For cell sorting, the cells were resuspended in F10 media (Life Technology, Carlsbad, California) before sorting with BD FACS Aria II SORP (BD Biosciences, San Jose, CA).