Fig. 4

HD180 iBMECs show unique responses to oxidative, angiogenic, and osmotic stress. A Schematic illustration of disease- and therapeutic-relevant perturbations to the HD BBB. B–D HD180 iBMECs are more vulnerable to oxidative damage: B time course of iBMEC TEER in response to various H₂O₂ concentrations. Red box denotes concentration resulting in most unique responses between cell sources. C iBMEC TEER 24 h after exposure to 0.6 mM H₂O₂. Data collected across n = 7 (HD180) and 6 (HD-corrected) independent differentiations. D Representative fluorescence images of cellular reactive oxygen species, nuclei, and f-actin (Phalloidin) 24 h after exposure to 0.6 mM. Red arrows indicate holes in endothelium. E Representative immunofluorescence images of VEGFR2. Data collected across n = 6 (HD180) and 4 (HD-corrected) independent differentiations. Quantification shown in Additional file 2: Fig. S2C. F Bead angiogenesis assay. Beads coated in iBMECs seeded in 6 mg mL collagen I + Matrigel, then supplemented with basal media or with 20 ng mL⁻¹ bFGF and 50 ng mL⁻¹ VEGF. Representative images show beads 72 h after treatment, where red asterisks denote angiogenic sprouts. G Quantification of sprout density across bead angiogenesis assay conditions. Data collected across n = 4 (HD180) and 3 (HD-corrected) independent differentiations. H Changes in TEER in response to osmotic treatment (10 min exposure to 1.4 M mannitol). Data collected across n = 5 independent differentiations