3D in vitro blood-brain-barrier model for investigating barrier insults

Abstract

The blood-brain-barrier (BBB) prevents that harmful substances in the blood enter the brain, and barrier disruption has been associated with a variety of central-nervous-system diseases. In vitro BBB models enable to recapitulate the BBB behavior in a controlled environment to investigate how the barrier reacts to stress events and external insults. Here, we present a human-cell-based BBB platform with integrated, transparent electrodes to monitor barrier tightness in real time at high spatiotemporal resolution. The BBB model includes human cerebral endothelial cells and primary human pericytes and astrocytes in a three-dimensional arrangement within a pump-free, open microfluidic platform. With our platform, we demonstrate that oxygen-glucose deprivation (OGD), which mimics the characteristics of an ischemic insult, induces a rapid remodeling of the cellular actin structures and subsequent morphological changes in the endothelial cells. High-resolution live imaging showed the formation of large actin stress-fiber bundles in the endothelial layer during OGD application, which ultimately led to cell shrinkage and barrier breakage. Simultaneous electrical measurements showed a rapid decrease of the barrier electrical resistance before the appearance of the stress fibers, which indicates that the barrier function is compromised already before the appearance of drastic morphological changes. The results demonstrate that our BBB platform recapitulates the main barrier functions in vitro and can be used to investigate the rapid reorganization of the BBB upon application of external stimuli.