New Blood-Brain Barrier Models Using Primary Parkinson's Disease Rat Brain Endothelial Cells and Astrocytes for the Development of Central Nervous System Drug Delivery Systems.

Abstract

Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by motor system defects due to the loss of dopaminergic neurons. A significant contributor to the current limited therapeutic treatments for PD is the poor penetration of potential drugs through the blood-brain barrier (BBB). The BBB is a highly specialized neurovascular system that separates components of the circulating blood from neurons. There is a great need to develop in vitro BBB models that retain fundamental characteristics and reliably predict the permeability of drug candidates. BBB breakdown may initiate and/or contribute to neuronal dysfunction and loss in diseases such as PD. However, there is no in vitro BBB model that mimics the pathological state of PD. To construct in vitro BBB models for drug delivery systems in the developing central nervous system (CNS), we isolated high purity endothelial cells from both normal and PD rat brain microvessels. The primary rat endothelial cell cultures maintained the properties of their in vivo counterparts. We developed and characterized in vitro rat endothelial cell and C6 glial cell coculture BBB models. We further examined the morphological and functional integrity of the barriers. The in vitro coculture BBB models we established displayed the typical cytoarchitecture and cellular markers by immunofluorescence staining and electron microscopy, high transendothelial electrical resistance (>300 Ω cm2), and a low permeability value (<3 × 10-6 cm/s). Our new models can be used to study BBB dysfunctions in relation to the pathogenesis and progression of PD, as well as a screening tool to test candidate drugs for PD treatment.