Human iPSC-Derived Blood-Brain Barrier Models

Blood-brain barrier (BBB) models derived from human induced pluripotent stem cells (iPSCs) are emerging and are considered promising tools for studying the central nervous system (CNS) diseases and drug development. The BBB consists of brain microvascular endothelial cells, astrocytes, pericytes, and basement membrane to maintain the homeostasis of the CNS, protect neurons from toxins, and regulate the transportation of nutrients. Currently available BBB models suffer from issues such as incomplete reconstruction of in vivo microenvironment, cells from non-uniform sources and batches, lack of multi-cell co-culture, and inability to simulate physiological dynamic and pathological state, which limit their use in the study of disease mechanisms and prediction of drug permeability.

Creative Biolabs provides comprehensive biotechnology services for BBB-related diseases that include basic mechanism research and disease model development to drug screening and personalized therapeutic solutions. With cutting-edge stem cell and microfluidic chip technologies, Creative Biolabs enables researchers and pharmaceutical companies to speed up BBB and neurological disease research and development.

Understanding the Blood-Brain Barrier

The BBB is formed mainly by brain microvascular endothelial cells, astrocytes, pericytes, and the basement membrane. The function of the BBB is to maintain homeostasis in the CNS, protect the neurons from toxic substances, and allow the diffusion of nutrients such as glucose, oxygen, and water.

The BBB has physical barriers, which are formed by tight junction proteins that restrict the diffusion of water-soluble molecules and allow the diffusion of lipid-soluble small molecules. Endothelial cells also express transporters and receptor-mediated transport systems that actively or passively transport certain molecules. Astrocytes communicate with endothelial cells to regulate BBB permeability and integrity. Pericytes also stabilize the vascular structure and maintain the BBB impermeability.

BBB also has metabolic barriers that regulate the chemical balance in the brain by actively transporting drugs or toxins out of the brain tissue via active transport systems. BBB is also related to ischemic stroke, neurodegenerative diseases, and neuroinflammation.

Figure 1 A diagram of BBB structure.^1,2

Construction of an iPSC-derived BBB Model

Differentiation Into Endothelial Cells

iPSCs are stimulated to differentiate into brain microvascular endothelial cells (BMECs) by using fully defined culture media (for example, Matrigel-coated) and signaling pathways (for example, Wnt/β-catenin, Notch). For example, BMEC-like cells with mature immune phenotypes can be generated by a specific method of endothelial cell culture (expanded endothelial cell culture methods or EECM). These cells have a morphology, barrier properties and endothelial adhesion molecule expression indistinguishable from primary human BMECs. Also, chemically defined media can increase the reproducibility and functionality of the differentiation.

Differentiation Into Astrocytes

NPCs are differentiated into astrocytes by using specific culture media (for example, culture media containing bFGF, EGF, CNTF and BMP4).

Co-Culture to Construct BBB Model

Endothelial cells, astrocytes, pericytes, and neurons are differentiated and co-cultured to reproduce complex BBB functions. For example, a microfluidic chip platform is used to study interactions between microglia and astrocytes. In this case, the chemokine ADP was found to enhance the migration of microglia. Also, Transwell membranes and a co-culture system are used to assess endothelial cell barrier function and the expression of transporter proteins.

iPSC-derived BBB Model Advantages

Table 1 Comparison of different BBB models

Functional Validation of the iPSC-BBB Model

Barrier integrity testing primarily involves the following methods:

• Immunofluorescence staining: used to locate the distribution of tight junction proteins, suitable for observing cell or tissue samples.
• Western Blot: Used to quantitatively analyze the expression levels of tight junction proteins, commonly used to detect proteins such as ZO-1, Occludin, and Claudin.
• Immunohistochemistry (IHC): Used for qualitative or semi-quantitative analysis of tight junction proteins in tissue samples.
• Trans-epithelial electrical resistance measurement (TEER): Used to assess the integrity of the epithelial cell layer, indirectly reflecting the function of tight junction proteins.

Applications of iPSC-BBB Models in Drug Permeability Studies

Applications of the iPSC-BBB model in drug permeability research can be seen in several aspects, including drug screening and development, disease model construction, transporter and drug interaction studies, integration of microfluidic chip technology, and integration with computational models. These studies provide new tools and methods for the treatment of central nervous system diseases.

Drug Permeability Prediction

The application of the iPSC-BBB model in drug permeability research can be used to predict drug permeability. A study based on iPSC-BBB models reported that the model could predict drug permeability with high accuracy, and it used the model to predict the permeability of 8 compounds. The predicted results were found to be highly consistent with in vivo permeability data (R2 = 0.83; P = 0.008). This suggests that iPSC-BBB models can be used as an effective tool for drug screening.

Drug Screening and Development

The iPSC-BBB model can also be applied to drug screening and development, helping researchers to identify drugs with potential therapeutic value. For example, one study used the iPSC-BBB model to evaluate the permeability and toxicity of multiple drugs and found that the model was effective in distinguishing CNS drugs from non-CNS drugs.

Disease Modeling and Personalized Therapy

The iPSC-BBB model can also be used for disease modeling to study the effects of specific diseases (such as Alzheimer's disease, Parkinson's disease, etc.) on the BBB. In addition, the model can also be used for personalized therapy by predicting the permeability of drugs in individuals based on patient-specific iPSC cells.

Transporter And Drug Interaction Studies

The iPSC-BBB model can also be applied to study the interactions between drugs and transporters, such as studying the efflux activity of P-glycoprotein (P-gp). Studies have shown that iPSC-BBB models can mimic the function of P-gp and be used to evaluate drug efflux activity.

Application of Microfluidic Chip Technology

Microfluidic chip technology can also be applied to construct iPSC-BBB models, simulating the BBB environment in vivo. For example, a study reported that it constructed a microfluidic chip that could maintain high TEER and mimic drug permeability at the level of in vivo.

Integration With Computational Models

The iPSC-BBB model can be combined with computational models to improve predictive performance. A study integrated molecular dynamics simulations and QSAR models to predict drug permeability and found that the two methods effectively complement the limitations of the iPSC-BBB model.

Why Creative Biolabs?

• Advanced Technology: Utilizes cutting-edge platforms like iPSC-derived BBB models and microfluidic chips for highly accurate and human-relevant research.
• Comprehensive Customization: Offers tailored solutions for different research needs, from cell line development to drug screening.
• Expert Team: Experienced scientists provide professional support and ensure high-quality, reproducible results.
• Efficiency and Reliability: Streamlined processes accelerate research timelines and reduce costs, making them a trusted partner for neuroscience and drug development.

Contact us today to discover how Creative Biolabs can support your research and accelerate your scientific breakthroughs.

References

1. Alahmari, Abeer. "Blood-Brain Barrier Overview: Structural and Functional Correlation." Neural Plasticity, edited by Long-Jun Wu, vol. 2021, Dec. 2021, pp. 1–10. DOI.org, https://doi.org/10.1155/2021/6564585.
2. Distributed under Open Access license CC BY 4.0, without modification.

Created June 2025

For Research Use Only. Not For Clinical Use.