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Setting-up an In Vitro Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated Transport
Published on: June 28, 2014
Membrane configuration optimization for a murine in vitro blood-brain barrier model
Diane M Wuest1, Allison M Wing, Kelvin H Lee
1Chemical and Biomolecular Engineering and Delaware Biotechnology Institute, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA.
Journal of Neuroscience Methods
|November 8, 2012
Summary
Optimizing membrane configuration in vitro enhances blood-brain barrier models. Smaller pores and PET membranes create tighter endothelial cell monolayers for better research.
Area of Science:
- Neuroscience
- Biomedical Engineering
- Cell Biology
Background:
- In vitro models of the blood-brain barrier (BBB) are crucial for studying its dynamic functions.
- Variability in model configurations hinders data comparison and understanding.
- Standardizing BBB in vitro models requires optimizing key parameters like membrane characteristics.
Purpose of the Study:
- To conduct a comprehensive screening study optimizing membrane configurations for in vitro BBB models.
- To identify influential membrane properties affecting cerebral endothelial cell monolayer tightness.
- To provide a standardized methodology for improved BBB model development.
Main Methods:
- Co-culture of primary murine brain endothelial cells and astrocytes on membranes with varying pore sizes (0.4–8.0 μm).
- Comparison of polyethylene terephthalate (PET) and polycarbonate (PC) membranes from different commercial sources.
- Characterization using transendothelial electrical resistance (TEER), sodium fluorescein permeability, and tight junction protein analysis.
Main Results:
- Non-contact co-culture orientation and smallest pore diameter (0.4 μm) significantly enhanced monolayer tightness.
- PET membranes from two manufacturers yielded significantly tighter monolayers compared to polycarbonate.
- Results were corroborated using the bEnd.3 murine brain endothelial cell line.
Conclusions:
- Membrane pore size and material significantly impact the formation of tight endothelial monolayers in vitro.
- Optimized membrane configurations using PET and smaller pore sizes advance BBB model development.
- The presented methodology offers a scalable approach for high-throughput BBB research.
