A homogenous cell population can be detected in FACS-analyses and ac-LDL-uptake assays one week after sorting. (a) Cells were grown in EGM2 MV for 1 week after sorting and analyzed with a light microscope for their phenotypic appearance. The cells were further analyzed by Flow Cytometry for expression of endothelial cell-specific surface markers CD31 (b), CD146 (c) and VEGF-R2 (d). Grey histograms indicate fluorescence signals of negative controls; white histograms indicate fluorescence signals of specific antigens. Results are representative of 4 separate experiments. Cells were incubated with 2.5 μg/ml ac-LDL-DiI for 4 h at 37°C and 5% CO2. Cells were subsequently washed twice with PBS and incubated with 5 μg/ml FITC-UEA for 1 h at 37°C and 5% CO2. After an additional washing step with PBS cells were analyzed by fluorescent microscopy (e: brightfield; f: fluorescence overlay). The scale bar depicts 100 μm.
Brandl et al. BMC Cell Biology 2014 15:12 doi:10.1186/1471-2121-15-12