Blood–brain-barrier organoids for investigating the permeability of CNS therapeutics
Name
nihms-1042217.pdf
Description
Accepted version
Size
722.85 KB
Format
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Checksum (MD5)
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Author(s)
Bergmann, Sonja
Lawler, Sean E.
Qu, Yuan
Fadzen, Colin M.
Wolfe, Justin
Regan, Michael S.
Pentelute, Bradley L.
Agar, Nathalie Y. R.
Cho, Choi-Fong
Date Issued
October 31, 2018
Journal
Nature Protocols
Publisher
Springer Nature
Citation
Bergmann, Sonja et al. "Blood–brain-barrier organoids for investigating the permeability of CNS therapeutics." Nature Protocols 13, 12 (December 2018): 2827–2843 © 2018 Springer Nature
Version
Author's final manuscript
Abstract
In vitro models of the blood–brain barrier (BBB) are critical tools for the study of BBB transport and the development of drugs that can reach the CNS. Brain endothelial cells grown in culture are often used to model the BBB; however, it is challenging to maintain reproducible BBB properties and function. ‘BBB organoids’ are obtained following coculture of endothelial cells, pericytes and astrocytes under low-adhesion conditions. These organoids reproduce many features of the BBB, including the expression of tight junctions, molecular transporters and drug efflux pumps, and hence can be used to model drug transport across the BBB. This protocol provides a comprehensive description of the techniques required to culture and maintain BBB organoids. We also describe two separate detection approaches that can be used to analyze drug penetration into the organoids: confocal fluorescence microscopy and mass spectrometry imaging. Using our protocol, BBB organoids can be established within 2–3 d. An additional day is required to analyze drug permeability. The BBB organoid platform represents an accurate, versatile and cost-effective in vitro tool. It can easily be scaled to a high-throughput format, offering a tool for BBB modeling that could accelerate therapeutic discovery for the treatment of various neuropathologies. Keywords: biological models; blood–brain barrier; cytological techniques; drug screening
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/s41596-018-0066-x
