title: An isogenic blood-brain barrier model comprising brain endothelial cells, astrocytes, and neurons derived from human induced pluripotent stem cells.
abstract: 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.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Scott G
authors.identifier: 0000-0003-1328-7365
authors.initials: SG
authors.lastname: Canfield
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Stebbins
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Bethsymarie Soto
authors.initials: BS
authors.lastname: Morales
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Shusaku W
authors.initials: SW
authors.lastname: Asai
authors.affiliation: Cedars-Sinai Medical Center, Board of Governors Regenerative Medicine Institute, Los Angeles, California, USA.
authors.forename: Gad D
authors.initials: GD
authors.lastname: Vatine
authors.affiliation: Cedars-Sinai Medical Center, Board of Governors Regenerative Medicine Institute, Los Angeles, California, USA.
authors.forename: Clive N
authors.initials: CN
authors.lastname: Svendsen
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Palecek
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
authors.forename: Eric V
authors.initials: EV
authors.lastname: Shusta
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D001812:Blood-Brain Barrier / Q000166:cytology / Q000502:physiology*; D001921:Brain / Q000166:cytology / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D008297:Male; D051379:Mice; D009474:Neurons / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
keywords: astrocytes; blood-brain barrier model; neurons; neurovascular unit; stem cells