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Flow cytometry

YR Ye Ran
YD Yankai Dong
YL Yuejiao Li
JX Jundong Xie
SZ Shubin Zeng
CL Chuanlei Liang
WD Wei Dai
WT Wenjing Tang
YW Yaojiong Wu
SY Shengyuan Yu
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The expression levels of cell surface proteins were determined by flow cytometry. Adherent cells were harvested by trypsin/EDTA (0.05%/0.02%), centrifuged, washed and resuspended in phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA). In order to evaluate the expression of intercellular adhesion in MSCs, aliquots of 1.0 × 106 cells were incubated with phycoerythrin (PE)-conjugated antibodies, CD49d (12–0499-73, eBioscience), CD106 (561679, BD Pharmingen) or PE antihuman CD324 (E-Cadherin; 324105, BioLegend) and corresponding isotype controls for 30 min before washing with PBS. Antibodies for MSC characterization were all purchased from BioLegend, which were fluorescein isothiocyanate (FITC)-conjugated monoclonal antibodies against CD45 (304005), CD34 (343503) and HLA-DR (307603) and allophycocyanin (APC)-conjugated antibodies against CD73 (344005), CD105 (endoglin; 323207) and CD90 (328113). All samples were analyzed by an EPICS XL flow cytometer (Beckman Coulter GmbH, Krefeld, Germany).

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