Fitc conjugated anti cd105

Cells were suspended in PBS containing 0.5% FBS and 2 mM EDTA at (1.0 × 10⁵) and incubated with phycoerythrin (PE)-conjugated anti-BST1 (clone: eBioSY11B5, eBioscience, 12-1579-4), fluorescent isothiocyanate (FITC)-conjugated anti-CD271 (clone: ME204, BioLegend, 345103), FITC-conjugated anti-CD90 (clone: 5E10, BioLegend, 328107), PE-conjugated anti-CD106 (clone: STA, BioLegend, 305805), FITC-conjugated anti-Stro-1 (BioLegend, 340105), PE-conjugated anti-MSCA-1 (clone: W8B2, BioLegend, 327305), FITC-conjugated anti-CD105 (clone: 43A3, BioLegend, 323203), FITC-conjugated anti-CD73 (clone: AD2, BioLegend, 344015), or PE-conjugated anti-CD146 (clone: SHM-57, BioLegend, 342003) antibody for 1 h at 4°C in the dark. Acquisition was performed with an EPICS XL ADC System (Beckman Coulter).

Cells were trypsinized and resuspended in washing buffer consisting of PBS with 2.5% FBS, and stained with primary antibodies for 30 min at 4 °C. Cells were then stained with streptavidin-PE/Cy5 (1 : 200; BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min at 4 °C in the dark. Primary antibodies used for cell staining were PE-conjugated anti-CD56 (1 : 20; Miltenyi Biotec), biotinylated anti-PDGFRα (2.5 μg/ml; R&D; cat. no. BAF322), FITC-conjugated anti-CD34 (1 : 10; BD Pharmingen), FITC-conjugated anti-CD45 (1 : 10; BD Pharmingen), FITC-conjugated anti-CD90 (1 : 200; BD Pharmingen), FITC-conjugated anti-CD105 (1 : 10; BioLegend, San Diego, CA, USA), and FITC-conjugated anti-CD166 (1 : 20; MBL, Aichi, Japan). Stained cells were analyzed by FACSCalibur or FACSVantage SE (BD Biosciences). Cell sorting was performed on a FACSVantage SE.