In Vitro Endothelial Cell Proliferation Assay Reveals Distinct Levels of Proangiogenic Cytokines Characterizing Sera of Healthy Subjects and of Patients with Heart Failure
Figure 1
In vitro endothelial cell proliferation in response to serum samples from healthy subjects. Primary HUVEC cultures, characterized by multiparametric flow cytometry analysis and morphological assessment (a), were used as target for in vitro proliferation assays performed by dynamic monitoring of cell proliferation using the xCELLigence system, in response to human serum samples (b)–(d). In (a), a representative field of a HUVEC monolayer observed by light microscopy is shown together with a representative multiparametric flow cytometry panel, displayed as dot plots, documenting the purity of the endothelial culture. In (b), a representative profile of HUVEC proliferation in the presence of EGM-2 complete medium and after removal of growth factors and serum (arrow) is shown. In (c), two representative profiles of endothelial cell proliferation in response to serum (20%) of an old healthy donor and a young healthy subject are shown. In (b) and (c), endothelial cell proliferation is shown as cell index (mean ± SD of samples assayed in quadruplicate) after normalization to the last cell index recorded before either depletion of medium serum and growth factors (in (b), arrow) or addition of human serum samples (in (c), arrow). In (d), HUVEC proliferation in response to human serum samples obtained from old and young healthy subjects was monitored up to 48 hours and expressed as normalized cell index (mean ± SD) calculated at the indicated time points. * compared to old healthy subjects at the corresponding time point.
