A Novel Transwell Blood Brain Barrier Model Using Primary Human Cells
Open accessAbstract
Structural alterations and breakdown of the blood brain barrier (BBB) is often a primary or secondary consequence of disease, resulting in brain oedema and the transport of unwanted substances into the brain. It is critical that effective in vitro models are developed to model the in vivo environment to aid in clinically relevant research, especially regarding drug screening and permeability studies. Our novel model uses only primary human cells and includes four of the key cells of the BBB: astrocytes, pericytes, brain microvascular endothelial cells (HBMEC) and neurons. We show that using a larger membrane pore size (3.0 µM) there is an improved connection between the endothelial cells, astrocytes and pericytes. Compared to a two and three cell model, we show that when neurons are added to HBMECs, astrocytes and pericytes, BBB integrity was more sensitive to oxygen-glucose deprivation evidenced by increased permeability and markers of cell damage. Our data also show that a four cell model responds faster to the barrier tightening effects of glucocorticoid dexamethasone, when compared to a two cell and three cell model. These data highlight the important role that neurons play in response to ischaemia, particularly how they contribute to BBB maintenance and breakdown. We consider that this model is more representative of the interactions at the neurovascular unit than other transwell models and is a useful method to study BBB physiology.
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BBB transwell models allow transport measurements but are low-throughput and do not capture transfection.
“BBB transwell models have a semipermeable membrane separating two compartments allowing for transport measurements but are low-throughput as they are typically only used in 12- or 24-well formats and do not capture transfection.30,31”
From Predictive High-Throughput Platform for Dual Screening of mRNA Lipid Nanoparticle Blood–Brain Barrier Transfection and Crossing (2024) · cites this paper as [31]
The Introduction says transwell models provide access to apical and basolateral compartments for drug application and sampling, supporting the transport-measurement point. The Methods compare 12- and 24-well setups, but do not establish that transwell models are typically limited to those formats or are low-throughput. No passage found addresses transfection. (LLM, conf=0.86)
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“Adding neurons to the culture makes the model more representative of the interaction in the NVU and increases the sensitivity of the BBB integrity to oxygen-glucose deprivation [115].”
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- Authors
- Stone, N. L., England, T. J., & O’Sullivan, S. E.
- Affiliations
- University of Nottingham (GB)
- Corresponding author
- Stone, Nicole L.
- Venue
- Frontiers in Cellular Neuroscience
- Publisher
- Frontiers Media SA
- Published
- 2019-06-06
- Volume
- 13
- Pages
- 230-230
- ISSN
- 1662-5102 (ISSN-L)
- PMC
- PMC6563620
- OpenAlex
- W2950636079
- Cited by
- 251
- References
- 5
- Citations by year
- 2026: 26 · 2025: 35 · 2024: 37 · 2023: 48 · 2022: 52 · 2021: 42 · 2020: 10 · 2019: 1
- FWCI
- 11.56
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Frontiers in Cellular Neuroscience (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Repository@Nottingham (University of Nottingham) (publishedVersion, cc-by)
- Repository@Nottingham (University of Nottingham) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Biotechnology and Biological Sciences Research Council
- Topics
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
- Keywords
- blood-brain barrier, transwell model, primary human cells, astrocytes, pericytes, brain microvascular endothelial cells, neurons, neurovascular unit, oxygen-glucose deprivation, ischemia, dexamethasone, blood-brain barrier permeability, brain oedema, drug screening
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