An isogenic blood–brain barrier model comprising brain endothelial cells, astrocytes, and neurons derived from human induced pluripotent stem cells

• Published in: Journal of neurochemistry 2017-03, Vol.140 (6), p.874-888
• Main Authors: Canfield, Scott G., Stebbins, Matthew J., Morales, Bethsymarie Soto, Asai, Shusaku W., Vatine, Gad D., Svendsen, Clive N., Palecek, Sean P., Shusta, Eric V.
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Animals; astrocytes; Astrocytes - physiology; Blood-brain barrier; Blood-Brain Barrier - cytology; Blood-Brain Barrier - physiology; blood–brain barrier model; Brain - cytology; Brain - physiology; Cell Differentiation - physiology; Cells, Cultured; Coculture Techniques; Endothelial Cells - physiology; Endothelium, Vascular - cytology; Endothelium, Vascular - physiology; Humans; Induced Pluripotent Stem Cells - physiology; Male; Mice; Neurons; Neurons - physiology; neurovascular unit; Rats; Rats, Sprague-Dawley; Stem cells
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/jnc.13923

The blood–brain barrier (BBB) is critical in maintaining a physical and metabolic barrier between the blood and the brain. The BBB consists of brain microvascular endothelial cells (BMECs) that line the brain vasculature and combine with astrocytes, neurons and pericytes to form the neurovascular unit. We hypothesized that astrocytes and neurons generated from human‐induced pluripotent stem cells (iPSCs) could induce BBB phenotypes in iPSC‐derived BMECs, creating a robust multicellular human BBB model. To this end, iPSCs were used to form neural progenitor‐like EZ‐spheres, which were in turn differentiated to neurons and astrocytes, enabling facile neural cell generation. The iPSC‐derived astrocytes and neurons induced barrier tightening in primary rat BMECs indicating their BBB inductive capacity. When co‐cultured with human iPSC‐derived BMECs, the iPSC‐derived neurons and astrocytes significantly elevated trans‐endothelial electrical resistance, reduced passive permeability, and improved tight junction continuity in the BMEC cell population, while p‐glycoprotein efflux transporter activity was unchanged. A physiologically relevant neural cell mixture of one neuron: three astrocytes yielded optimal BMEC induction properties. Finally, an isogenic multicellular BBB model was successfully demonstrated employing BMECs, astrocytes, and neurons from the same donor iPSC source. It is anticipated that such an isogenic facsimile of the human BBB could have applications in furthering understanding the cellular interplay of the neurovascular unit in both healthy and diseased humans. Read the Editorial Highlight for this article on page 843. We developed an isogenic human blood–brain barrier (BBB) model comprising induced pluripotent stem cell (iPSC) derived brain endothelial cells (BMECs), astrocytes, and neurons. We have demonstrated that iPSC‐derived neurons and astrocytes can improve the tight junctions and induce barrier tightening in iPSC‐derived BMECs. An isogenic BBB model where each cell type is derived from the same patient iPSC source would enable new approaches in brain disease modeling and drug development. Read the Editorial Highlight for this article on page 843.