Abstract
Constructing a reliable in vitro blood–brain barrier (BBB) model using human primary cells has been considered a major challenge during the past decades. These systems could provide valuable information regarding the effect of therapeutic compounds on different BBB cell types (endothelial cells, astrocytes, pericytes) and their ability to cross the barrier in order to reach the brain. Several attempts have been made to develop in vitro BBB models, but these studies mainly used rat, bovine, and porcine cells rather than human primary cells. Genetically modified cell lines have also been used, but they do not appear to maintain physiological properties of the BBB. Here, we describe a detailed protocol for co-culturing and maintaining human brain primary endothelial cells, pericytes, and astrocytes under flow to create an in vitro human BBB model, which can be used for toxicity testing and for studying cross-interaction among different cell types involved in the BBB formation.
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Acknowledgements
This work was supported by the funding from the Biotechnology and Biological Sciences Research Council (BBSRC) and the Innovate UK. We acknowledge the support from Dr Kelly Davidge and Dr Malcolm Wilkinson at Kirkstall Ltd. for technical help with the QV500 system used in this study.
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Miranda-Azpiazu, P., Saha, S. (2022). A Novel Dynamic Human In Vitro Model for Studying the Blood–Brain Barrier. In: Stone, N. (eds) The Blood-Brain Barrier. Methods in Molecular Biology, vol 2492. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2289-6_9
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DOI: https://doi.org/10.1007/978-1-0716-2289-6_9
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