Fig. 3
Functional characterization of BBB properties of mBECs. A Transendothelial electrical resistance (TEER, Ω cm2) of confluent mBEC monolayers on Collagen IV/Fibronectin coated 1 µm pore transwell inserts cultured in the presence or absence of 10 µM all-trans Retinoic Acid (± RA) and ACM in the abluminal chamber (mean ± SD) assessed over 11 days in culture. Results are representative of 3 independent differentiations. B Comparison of effect of transwell seeding densities on TEER values (mean ± SD). C Comparison of laminin matrix coatings: Laminin 511 (511), Laminin 521 (521), Laminin 211 (211), and Collagen IV/Fibronectin (C/F) on TEER induction (mean ± SD) over a 7 day period. Results are representative of 3 independent differentiations. D Comparison between TEER values (left y-axis) and sodium fluorescein permeability coefficient (Pe, right y-axis) in mBECs from 6 independent differentiations during protocol optimization (x-axis). The means reflect variability from different differentiations during protocol optimization steps (mean ± SD). E Comparison between TEER values (left y-axis) and sodium fluorescein permeability coefficient (Pe, right y-axis) in mBECs, human iPSC-derived BECs (iBECs), bEnd.3, SV-ARBECs and pmBECs (mean ± SD). Results are representative of 6 independent differentiations. F Permeability values of Rhodamine123 from apical to basolateral (A-B) and basolateral to apical (B-A) compartments. Efflux ratio (B-A/A-B) for Rhodamine is 1.94 (mean ± SD). Results are from 2 independent differentiations