(1004-A) A perfused 3D model of the human blood-brain barrier in the OrganoPlate®

Abstract: The vasculature of the brain is made up of highly specialized endothelial cells that form a tight blood–brain barrier (BBB). The BBB restricts the entry of harmful circulating molecules into the brain and ensures a homeostatic microenvironment for healthy brain functioning. While the BBB protects the brain, its selective barrier properties can also complicate drug delivery to the brain for treatment of various central nervous system (CNS) disorders. Better models of the human BBB can help increase our understanding of BBB function, assess potential toxicity of novel compounds, and improve drug delivery technologies.
We here report a perfused 3-dimensional (3D) model of the human BBB in a high-throughput microfluidic platform that allows culture of up to 64 independent BBBs on-chips. The model comprises a lumenized vessel of primary human brain microvascular endothelial cells (HBMECs) grown against an extracellular matrix gel. Immunofluorescence staining confirmed expression of endothelial adherens and tight junction proteins. The model demonstrates tight barrier function as evidenced by retention of small molecule sodium fluorescein in its lumen. Exposure to the toxic compound staurosporine led to a concentration-dependent disruption of the barriers resulting in reduced transendothelial electrical resistance (TEER) and increased permeability to sodium fluorescein. Furthermore, expression of relevant BBB transporters was confirmed in the model by PCR and function of P-glycoprotein (P-gp) was strongly inhibited in presence of a P-gp inhibitor. The BBB on-a-chip model presented here can be used for fundamental studies of BBB function in health and disease and for investigation of potential restorative therapies to fight neurological disorders. Due to the platform’s relatively high throughput and compatibility with automation, the model holds potential for drug compound screening.