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Fig. 5 | Fluids and Barriers of the CNS

Fig. 5

From: Application of blood brain barrier models in pre-clinical assessment of glioblastoma-targeting CAR-T based immunotherapies

Fig. 5

CAR-F263 extravasation and U87vIII cytotoxicity in BTBB-on-chip model. A Schematic illustrating sequence of immune cell adhesion and extravasation across the BEC monolayers in the presence of shear stress. B Image of CAR-F263, labeled with CytoLight Rapid Green Reagent, extravasation through the pores of the SynBBB chips. Arrow: CAR-F263 traversing through 3 µm pore. C–D Visualization of CAR-F263 in the of U87vIII channel post-extravasation. Scale bar = 50 µm (E) Representative images of U87vIII-mKate2 cells (red), pre (0 h) and post (24 and 48 h) perfusion of CAR-F263. U87vIII-mKate2 cell death is observed at 48 h post-CAR-F263 perfusion indicating post BBB cytotoxicity. Scale bar = 100 µm. F Perfusion of the middle iBEC channel with sodium fluorescein (NaFl–green) confirming intact BBB barrier integrity (before CAR-F263 perfusion) and evidence of barrier disruption (leakiness) after CAR-F263 perfusion. Evidence of BBB disruption is visualized by NaFl signal leaking into the adjacent channels. G Assessment of barrier integrity by quantification of average NaFl ratio intensity in the outer to inner channels using ImageJ (mean + SD)

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