Characterization of a Primate Blood-Brain Barrier Co-Culture Model Prepared from Primary Brain Endothelial Cells, Pericytes and Astrocytes.

Daisuke Watanabe,Andrea E Tóth,Shinsuke Nakagawa,Jun Aruga,Masami Niwa,Yoichi Morofuji,Monika Vastag,Mária A Deli

doi:10.3390/pharmaceutics13091484

Daisuke Watanabe, Andrea E Tóth + Show 6 more

Open Access

https://doi.org/10.3390/pharmaceutics13091484

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Abstract

Culture models of the blood-brain barrier (BBB) are important research tools. Their role in the preclinical phase of drug development to estimate the permeability for potential neuropharmaceuticals is especially relevant. Since species differences in BBB transport systems exist, primate models are considered as predictive for drug transport to brain in humans. Based on our previous expertise we have developed and characterized a non-human primate co-culture BBB model using primary cultures of monkey brain endothelial cells, rat brain pericytes, and rat astrocytes. Monkey brain endothelial cells in the presence of both pericytes and astrocytes (EPA model) expressed enhanced barrier properties and increased levels of tight junction proteins occludin, claudin-5, and ZO-1. Co-culture conditions also elevated the expression of key BBB influx and efflux transporters, including glucose transporter-1, MFSD2A, ABCB1, and ABCG2. The correlation between the endothelial permeability coefficients of 10 well known drugs was higher (R2 = 0.8788) when the monkey and rat BBB culture models were compared than when the monkey culture model was compared to mouse in vivo data (R2 = 0.6619), hinting at transporter differences. The applicability of the new non-human primate model in drug discovery has been proven in several studies.

Highlights

We have demonstrated that astrocytes, but brain pericytes can increase the tightness of the paracellular barrier of rat brain endothelial cells in culture [9]
Our results show that astrocytes, pericytes, and their combination do not uniformly increase the level of different influx and efflux transporters, but a complex pattern of regulation is seen, which is in agreement with findings on porcine brain endothelial models [26]
We have developed and characterized a non-human primate co-culture blood-brain barrier (BBB) model using primary monkey brain endothelial cells, rat brain pericytes, and astrocytes

Summary

Introduction

Cell-culture based models have greatly contributed to our knowledge on the physiology, pathology, and pharmacology of the blood-brain barrier (BBB) [1,2,3]. In the preclinical phase of drug development, it is key to estimate the brain penetration of potential neuropharmaceuticals, and in this process culture models of the BBB are essential tools [2,4,5,6]. The specific features of cerebral capillary endothelial cells, which form the anatomical basis of the BBB, are organ specific. The dynamic interactions between brain endothelial cells and the neighboring astrocytes and pericytes promote the development and maintenance of BBB functions [7,8]. It is crucial to mimic this cellular complexity and the crosstalk

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