Cd105 560839

Flow cytometry was performed to characterize hAD-MSCs. The following phycolerythrin (PE)-labeled antibodies were used to analyze cell phenotypes: anti-human CD29 (557332), CD31 (560983), CD34 (550761), CD44 (562818), CD45 (560975), CD73 (562430), CD90 (561970), and CD105 (560839) (all diluted 1：10; BD Co., San Diego, CA, USA). Next, 1×10⁶ cells were stained with 10 μL each antibody in 100 μL phosphate-buffered saline (PBS) for 15 min at room temperature. Flow cytometry was performed using the Guava easyCyte 6HT (EMD Millipore, Billerica, MA, USA), and the data were examined with Guava Incyte software (version 2.8, EMD Millipore).

Immunophenotyping of eMSCs [cluster of differentiation (CD) marker expression] was performed using an Epics XL flow cytometer (Beckman Coulter, Inc., Brea, CA, USA). A single cell suspension was obtained using 0.05% trypsin/EDTA. Cells (1×10⁶) were suspended in 1 ml PBS containing 5% FCS. Subsequently, according to the manufacturer's instructions, the cells were incubated at room temperature for 30 min with the following mouse anti-human fluorescein isothiocyanate-conjugated monoclonal antibodies: anti-CD34 (555821; 1:250), CD45 (555482; 1:250) and CD90 (555595; 1:250), and phycoerythrin-conjugated monoclonal antibodies: Anti-CD19 (557835; 1:250), CD73 (561014; 1:200), CD105 (560839; 1:250), CD146 (561013; 1:200) and human leukocyte antigen (HLA)-DR (555812; 1:200; all BD Pharmingen, San Diego, CA, USA). All antibodies were pre-diluted for use according to the recommended volume per test. For each test, 1×10⁶ cells in 100 µl PBS were used. For CD73, CD146 and (HLA)-DR the volume per test was 20 µl. Cells were analyzed by flow cytometry.