Cd90 fluorescein isothiocyanate fitc

In accordance with the appraisal standards of stem cells proposed by the International Society for Cellular Therapy (ISCT), the NP-progenitor cell surface marker Tie2 and the MSC surface markers CD73, CD90, CD105, CD34, and CD45 were measured. Isotype controls (BD Biosciences, NJ, USA) were used in each case as the negative control. To put it simply, 2 × 10⁵ NPMSC were incubated with monoclonal primary antibodies (CD105-phycoerythrin (PE), CD90-fluorescein isothiocyanate (FITC), CD73-PE, CD45-FITC, CD34-PE, and Tie2-FITC (BD Biosciences, NJ, USA)) for 30 min at room temperature in the dark. Then, cells were washed twice with cold PBS and resuspended in 500 μL PBS to produce a single cell solution, and specific surface marker expression was detected using flow cytometry (FACSCalibur, BD Biosciences).

MSCs were harvested from culture flasks using trypsin-EDTA (as above) at 37°C and then washed twice with PBS. Next, cell suspensions were incubated with antibodies against CD44-phycoerythrin (PE), CD73-PE, CD14-PE, CD34-PE, CD90-fluorescein isothiocyanate (FITC), CD45-FITC, and CD105-PerCP, respectively (all from BD Biosciences, USA), and protected from light for 30 min at 4°C. After incubation, the cells were washed twice with PBS. Fluorescence intensity was measured using flow cytometry (Beckman Coulter FC500, USA) and the data were analyzed with CXP software (Beckman Coulter FC500).