Figure 4.

Reactive hPSC-derived astrocytes contribute to blood-brain barrier disruption

Co-cultures were performed using a transwell system, with brain microvascular endothelial cells (BMECs) seeded on the transwell and astrocytes seeded in the bottom of the well.

(A–I) Representative images of iPSC-derived BMECs expressing characteristic endothelial cell markers, including Claudin-5, glucose transporter 1 (GLUT-1), Occludin, and zonula occludens 1 (ZO-1). Reactive astrocytes induced a reduction in transendothelial electrical resistance (TEER) levels (G), increased fluorescein permeability (H), and elevated rhodamine 123 transport (I), compared to the effect promoted by control astrocytes. CsA, cyclosporin. ANOVA followed by Tukey’s multiple comparison test in (G) and (H) and by Šídák’s multiple comparisons test in (I). Data represent mean values ±SEM. ^∗p < 0.05 and ^∗∗p < 0.01, BMEC monoculture vs. BMECs co-cultured with control astrocytes; ^###p < 0.001, BMEC monoculture vs. BMECs co-cultured with reactive astrocytes; ^&&p < 0.01, BMECs co-cultured with control astrocytes vs. BMECs co-cultured with reactive astrocytes in (G). ^∗∗∗p < 0.001 and ^∗∗∗∗p < 0.0001 in (H) and (I). Scale bar: 50 μm.