Figure 7
Impact of GAG derivatives and role of VEGFR-2 for the pro-angiogenic effect of GAG derivatives on sprouting of HUVEC spheroids. VEGF165 was used as a positive control for spheroid sprouting in a 3D in vitro angiogenesis assay. (a) Sprouting of HUVEC spheroids determined by light microscopy. (b) Cumulative sprouting length calculated using ImageJ and (c) number of sprouts. For (a) one representative microscope picture is shown and for (b) and (c) values represent the mean ± SD of n = 3. One-way ANOVA: **p < 0.01; ***p < 0.001 vs. Ctrl. (d-f) Cells were treated with a VEGFR-2 neutralizing antibody (VEGFR-2 Ab) and GAG. (d) Sprouting of HUVEC spheroids determined by light microscopy. (e) Cumulative sprouting length calculated using ImageJ and (f) number of sprouts. For (d) one representative microscope picture is shown and for (e) and (f) values represent the mean ± SD of n = 3 with 10 spheroids evaluated for each independent experiment. One-way ANOVA: *p < 0.05; **p < 0.01; ***p < 0.001 vs. VEGF + VEGFR-2 Ab; ###p < 0.001 vs. Ctrl. [adapted and modified from33].
