Fig. 5

Serum-derived factors in CKD disrupted BBB integrity in vitro. A–C Effects of CKD dialysis serum on the iPSC-derived human brain microvascular endothelial cells (ihBMEC) monolayer. Incubation with CKD dialysis serum significantly decreased A TEER, and increased B tracer (sodium fluorescein) permeability across the ihBMEC monolayer 3 days after serum treatment initiation, compared with serum from healthy volunteers. C ihBMEC viability measured by CCK8 assay 3 days after serum treatment initiation was not changed. D, E Effects of uremic toxins on bEnd.3 endothelial barrier. D TEER remained significantly lower in the 3Tox + Urea + LPS group compared with the control group. After 4 days of toxin exposure, TEER was significantly lower in the Urea + LPS (p < 0.01), 3Tox + LPS (p < 0.05), and 3Tox + Urea + LPS group (p < 0.0001) compared with the control group. E bEnd.3 cell viability was assessed by trypan blue assay and was equivalent across groups. 3Tox = indoxyl sulfate 150 μg/mL + p-cresyl sulfate 100 μg/mL + trimethylamine N-oxide 1 mg/mL. The reference Cells Only group was cells grown in DMEM with 10% FBS, and mannitol 450 mg/dL was used as an osmotic control group. Data shown are mean ± SEM of at least 3 independent experiments done in duplicate. *p < 0.05 and **p < 0.01