Blood-Brain Barrier Model
Blood brain barrier (BBB) models are essential for studying the transport efficacy of drugs that target the brain, and for understanding the pathological neurovascular functions in various diseases. Despite the high complexity and high cost of in vivo animal BBB models, 80% of drug candidates that cleared the animal tests failed subsequently in the clinical trials. Therefore, a highly robust, predictive and cost-effective in vitro BBB model is needed. To address this issue, microfluidic technology is utilized to create a 3D BBB model through the co-culture of human induced pluripotent stem cell (iPSC)-derived endothelial cells (EC), brain pericytes (PC) and astrocytes (AC). These all-human cells self-organise into a microvascular network within a fibrin gel through vasculogenesis. The BBB model not only exhibits physiologically relevant structures such as tight junction proteins, but more importantly, its permeability is comparable to in vivo values measured in rat brain. This in vitro BBB model can therefore be used to screen for brain-targeting drugs or to study the neurovascular functions.
Protocol
This protocol is based on work by Campisi et al in reference (1). We thank Dr. Marco Campisi for his contribution to the preparation of this document.
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Key References
- A fully iPS-cell-derived 3D model of the human blood–brain barrier for exploring neurovascular disease mechanisms and therapeutic interventions. González-Gallego, J., Todorov-Völgyi, K., Müller, S. A., Antesberger, S., Todorov, M. I., Malik, R., … & Paquet, D. (2025). Nature Neuroscience, 1-14
- Self-assembling 3D vessel-on-chip model with hiPSC-derived astrocytes. Nahon, D. M., Cuenca, M. V., van den Hil, F. E., Hu, M., de Korte, T., Frimat, J. P., … & Orlova, V. V. (2024). Stem Cell Reports, 19(7), 946-956
- Unveiling the Influence of Tumor Microenvironment and Spatial Heterogeneity on Temozolomide Resistance in Glioblastoma Using an Advanced Human In Vitro Model of the Blood-Brain Barrier and Glioblastoma. Lam, M. S., Aw, J. J., Tan, D., Vijayakumar, R., Lim, H. Y. G., Yada, S., … & Pavesi, A. (2023). Small, 19(52), 2302280
- Simultaneous induction of vasculature and neuronal network formation on a chip reveals a dynamic interrelationship between cell types. Isosaari, L., Vuorenpää, H., Yrjänäinen, A., Kapucu, F. E., Kelloniemi, M., Pakarinen, T. K., … & Narkilahti, S. (2023). Cell Communication and Signaling, 21(1), 132
- Bioengineered perfused human brain microvascular networks enhance neural progenitor cell survival, neurogenesis, and maturation. Winkelman, M. A., & Dai, G. (2023). Science Advances, 9(19), eaaz9499
- Biosensors Integration in Blood–Brain Barrier-on-a-Chip: Emerging Platform for Monitoring Neurodegenerative Diseases. Mir, M., Palma-Florez, S., Lagunas, A., López-Martínez, M. J., & Samitier, J. (2022)
- Interstitial flow enhances the formation, connectivity, and function of 3D brain microvascular networks generated within a microfluidic device. Winkelman, M. A., Kim, D. Y., Kakarla, S., Grath, A., Silvia, N., & Dai, G. (2022).Lab on a Chip, 22(1), 170-192
- Engineered human blood–brain barrier microfluidic model for vascular permeability analyses. Hajal, C., Offeddu, G. S., Shin, Y., Zhang, S., Morozova, O., Hickman, D., … & Kamm, R. D. (2022). Nature protocols, 17(1), 95-128
- Engineered 3D vessel-on-chip using hiPSC-derived endothelial- and vascular smooth muscle cells. Cuenca, M. V., Cochrane, A., van den Hil, F. E., de Vries, A. A., Oberstein, S. A. L., Mummery, C. L., & Orlova, V. V. (2021). Stem cell reports, 16(9), 2159-2168
- Study of the neurotoxicity of indoor airborne nanoparticles based on a 3D human blood-brain barrier chip. Li, Y., Liu, Y., Hu, C., Chang, Q., Deng, Q., Yang, X., & Wu, Y. (2020). Environment international, 143, 105598
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