Figure 2
Functional characterization of monkey IPSC-ECs.
(A) Flow cytometry analysis of cIPSC-ECs at day 13 of differentiation. Cells express endothelial markers CD144, CD31 and CD309 and stain negative for hematopoietic lineage marker CD45. Unstained control samples are shown in grey. (B) Uptake of acetylated LDL by cIPSC-ECs. Overlay of phase contrast image and fluorescent channel shows incorporated acLDL (conjugated to Alexa488) in ECs. Scale bar: 50 μm. (C) Angiogenic potential of cIPSC-ECs demonstrated by tube formation assay. After 24 hours, cells form a network of tubular structures. Tube formation is inhibited in the presence of 10 μM sulforaphane or 2 μg/ml anti-VEGF antibody. Scale bars: 200 μm. Quantification of the inhibitory effect of anti-angiogenic molecules on the tubulogenesis of cIPSC-ECs (right panel). Columns show total tube length relative to control from three independent experiments. (D) Impedance-based monitoring of cIPSC-EC culture demonstrating the formation of a tight monolayer. One of 2 independent experiments performed is depicted (n = 4 technical replicates), error bar STD. (E) Response to proinflammatory stimuli. Twenty-four hours after stimulation with proinflammatory cytokines, cIPSC-ECs exhibit upregulated EC. activation marker ICAM1. Scale bars: 50 μm. Quantification of median intensity of ICAM1 staining (right panel). Columns show mean +/− STD of three independent experiments and data were analyzed using Student’s t-test.
