Miniaturization and Automation of a Human In Vitro Blood-Brain Barrier Model for the High-Throughput Screening of Compounds in the Early Stage of Drug Discovery.

Elisa L J Moya,Elodie Vandenhaute,Nathalie Maubon,Fabien Gosselet,Marie-Christine Boucau,Eleonora Rizzi,Marie-Pierre Dehouck,Johan Hachani

doi:10.3390/pharmaceutics13060892

Elisa L J Moya, Elodie Vandenhaute + Show 6 more

Open Access

https://doi.org/10.3390/pharmaceutics13060892

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Journal: Pharmaceutics	Publication Date: Jun 16, 2021
Citations: 12	License type: CC BY 4.0

Affiliation: HRA Pharma (France), Artois University

Abstract

Central nervous system (CNS) diseases are one of the top causes of death worldwide. As there is a difficulty of drug penetration into the brain due to the blood–brain barrier (BBB), many CNS drugs treatments fail in clinical trials. Hence, there is a need to develop effective CNS drugs following strategies for delivery to the brain by better selecting them as early as possible during the drug discovery process. The use of in vitro BBB models has proved useful to evaluate the impact of drugs/compounds toxicity, BBB permeation rates and molecular transport mechanisms within the brain cells in academic research and early-stage drug discovery. However, these studies that require biological material (animal brain or human cells) are time-consuming and involve costly amounts of materials and plastic wastes due to the format of the models. Hence, to adapt to the high yields needed in early-stage drug discoveries for compound screenings, a patented well-established human in vitro BBB model was miniaturized and automated into a 96-well format. This replicate met all the BBB model reliability criteria to get predictive results, allowing a significant reduction in biological materials, waste and a higher screening capacity for being extensively used during early-stage drug discovery studies.

Highlights

After two days of cell thawing and growing in petri dishes, noncontact cocultures were reproduced by firstly seeding bovine brain pericytes (PCs) on bottom-well plates coated with gelatine, using a basal endothelial cell medium (ECM; Sciencell, Carlsbad, CA, USA) supplemented with 5% fetal calf serum (FCS), 1% endothelial cell growth supplement (Sciencell, Carlsbad, CA, USA) and 0.5% gentamicin (Biochrom AG, Berlin, Germany), and placed for 3 h at 37 ◦ C
Endothelial cells derived from hematopoietic stem cells CD34+ and brain pericytes were thawed and let grown in petri dishes two days before the cells seeding in the TW, counting this step as day 0
For the first time in the field, the development of a miniaturized and automated in vitro human blood–brain barrier (BBB) model for the high-throughput screening systems (HTS) of the compounds in 96-multiwell systems has increased the facility of Central nervous system (CNS) drug/compound screenings

Summary

Introduction

There are still a large number of unmet medical needs concerning the treatment of most CNS diseases [3]. The development of drugs for treating CNS disorders is a medically challenging and commercially risky field. Numerous medical research efforts are focused on finding drug candidates for CNS disease treatments, but as the brain is one of the more protected organs, few neurological disorders have fully benefited from pharmacotherapy. One of the main limitations for the treatment of neurological disorders is the difficulty to deliver drugs to the brain. This protection mainly comes from the blood–brain barrier (BBB), which is a selective barrier formed by the endothelial cells of cerebral microvessels.

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