Production of Human Endothelial Cells Free from Soluble Xenogeneic Antigens for Bioartificial Small Diameter Vascular Graft Endothelization

Figure 1

hASCs characterization and endothelial differentiation. (a) Flow cytometry analysis of hASCs expression of mesenchymal stem cell markers. Black line indicates isotype-matched monoclonal antibody controls and red line indicates positive stained cells. (b) RT-PCR analysis of VEGFR2 and vWF mRNA expression during endothelial differentiation of hASCs. The expression of those markers was also assessed at protein level by immunohistochemistry (c). Green fluorescence corresponds to vWF protein presence (c1, c2, and c3), red corresponds to VEGFR protein expression (c4 and c5), and blue corresponds to cell nuclei (c1–c5). (d) Functional analysis of endothelial cell differentiation, as assessed by tube formation assay. In (d1) to (d3), inverted microscopy was used to document tube formation. In (d4) to (d6), cells were stained with calcein-AM and observed under fluorescence microscope, in order to show cell viability in addition to tube formation. for each experiment.

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