title: Improved method for the preparation of a human cell-based, contact model of the blood-brain barrier.
abstract: The blood-brain barrier (BBB) comprises impermeable but adaptable brain capillaries which tightly control the brain environment. Failure of the BBB has been implied in the etiology of many brain pathologies, creating a need for development of human in vitro BBB models to assist in clinically-relevant research. Among the numerous BBB models thus far described, a static (without flow), contact BBB model, where astrocytes and brain endothelial cells (BECs) are cocultured on the opposite sides of a porous membrane, emerged as a simplified yet authentic system to simulate the BBB with high throughput screening capacity. Nevertheless the generation of such model presents few technical challenges. Here, we describe a protocol for preparation of a contact human BBB model utilizing a novel combination of primary human BECs and immortalized human astrocytes. Specifically, we detail an innovative method for cell-seeding on inverted inserts as well as specify insert staining techniques and exemplify how we use our model for BBB-related research.
authors.affiliation: Australian Centre for Blood Diseases, Monash University.
authors.forename: Be'eri
authors.initials: B
authors.lastname: Niego
authors.forename: Robert L
authors.initials: RL
authors.lastname: Medcalf
mesh_terms: D001253:Astrocytes / Q000166:cytology*; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000502:physiology*; D001921:Brain / Q000098:blood supply*; D002461:Cell Line, Transformed; D018920:Coculture Techniques / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D008567:Membranes, Artificial
chemical_list: D008567:Membranes, Artificial