(1298-C) Generation of Human Endothelial Cells for Integration of Pericytes and Regional Specific Astrocytes to Mimic in vitro Blood Brain Barrier Model from Human Induced Pluripotent Stem Cells

Abstract: The Human blood brain barrier (BBB) is a protective microvascular system that encapsulates the central nervous system (CNS) and is crucial for maintaining a homeostatic environment for the brain. Breakdown of the BBB’s integrity is implicated in several neurodegenerative diseases: Alzheimer’s Disease (AD), Parkinson’s disease (PD), Amyotrophic lateral Sclerosis (ALS), Multiple Sclerosis (MS), and Hunting’s disease, to name a few. Unfortunately, availability of primary human BBB and its cellular components are limited. Thus, precise recapitulation of human BBB in vitro is an efficient tool for pharmaceutical research and applications. This study demonstrates efficient methods to differentiate human induced pluripotent stem cells (hiPSCs) into three cellular components to mimic human BBB in vitro. We differentiated, and identified each cell type by morphology and specific marker expression as follows: VE-cadherin+ZO1+Glut1+ endothelial cells (EC), PDGFRβ+NG2+ pericytes (PC) and GFAP+ astrocytes (AC). To characterize and assess barrier function of our human BBB in vitro, we optimized a tri-culture system with the hiPSC derived EC, PC, and AC, and measured the system’s transepithelial/ transendothelial electrical resistance (TEER) as well as permeability. Our TEER results indicate, the hiPSC-derived triculture of ECs, PCs, and ACs formed a monolayered which like a barrier. Because of its functional properties, the human BBB, is emerging as a critical target for pharmaceutical approaches. Therefore, hiPSC-based BBB modeling can be a reproducible strategy for disease modeling and drug screening in neurodegenerative diseases.