Fig. 2.

Endothelial cell cultures on crosslinked collagen films without peptide, functionalized with GFOGER, with GLOGEN, with VWFIII_Nleand with GFOGER + VWFIII_Nle, or non-crosslinked films. A) EC spreading. After 45 min at 37 °C with 5% CO₂, hESC-derived ECs were fixed and stained with Rhodamine-Phalloidin. Representative fields of view for GFOGER and GLOGEN are shown. Mean cell area was calculated from 10 randomly selected fields of view per conditions in three independent repeats (n = 3) and plotted as mean ± s.e.m. THP functionalization consistently increased mean cell surface area (one-way ANOVA, p < 0.001). Columns were compared to crosslinked films with no peptides using Dunnett's post-hoc test. B) EC proliferation. hESC-derived ECs were cultured for 24 h at 37 °C with 5% CO₂. EdU was introduced in the media and cells were left for 2h at 37 °C with 5% CO₂. Cells were fixed, nuclei were stained with Hoechst 33342 and EdU was stained with Alexa Fluor-488. The ratio of cells with EdU-Alexa488 over the total number of cells was calculated from 10 randomly selected fields of view per conditions in three independent repeats (n = 3) and plotted as mean ± s.e.m. THP functionalization consistently increased the percentage of proliferating ECs after 24h (one-way ANOVA, p < 0.01). Columns were compared to crosslinked films with no peptides using Dunnett's post-hoc test. C) EC LDL uptake. hESC-derived ECs were cultured in cell culture media containing Ac-LDL-Dil for 24 h at 37 °C with 5% CO₂, fixed and stained with Hoechst 33342. Representative fields of view are shown. Mean Dil fluorescence intensity per cell was measured in 10 randomly selected fields of view per conditions in four independent repeats (n = 4) and plotted as mean ± s.e.m. THP functionalization consistently increased the uptake of acetylated LDL by ECs (one-way ANOVA, p < 0.01). Columns were compared to crosslinked films with no peptides using Dunnett's post-hoc test.