Establishment of an In Vitro Model of Human Blood-Brain Barrier to Study the Impact of Ischemic Injury.

The blood-brain barrier (BBB), mainly composed of brain microvascular endothelial cells, astrocyte end-feet, and pericytes, serves as a physical and biochemical barrier that selectively limits the passage of circulating molecules into the brain parenchyma. The disruption of its integrity and function is a major cause of increased mortality and disability among ischemic stroke patients. Hence, scrutiny of the cellular and molecular mechanisms that alter BBB permeability following an ischemic injury remains of paramount importance. In this context, establishment of an in vitro model of BBB that closely simulates human cerebral barrier may offer an easy, inexpensive, and straightforward approach to identify signaling pathways involved in BBB breakdown and may help to discover new therapeutic targets to restore its damage. This chapter describes a sequential method pertaining to establishment of a triple culture model of human BBB consisting of the three main cellular components of the cerebral barrier. It also documents how the integrity and function of this barrier are evaluated through measurements of transendothelial electrical resistance (TEER) and paracellular flux of permeability marker and sodium fluorescein (NaF, 376 Da), respectively, both in normal and experimental conditions mimicking ischemic injury.