Generation of a Human iPSC-Based Blood-Brain Barrier Chip
| Autor: | Anna Herland, Michael J. Workman, Clive N. Svendsen, Gad D. Vatine, Srikanth Jagadeesan |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell type General Chemical Engineering Induced Pluripotent Stem Cells Models Neurological Central nervous system Blood–brain barrier General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Organ Culture Techniques 0302 clinical medicine Lab-On-A-Chip Devices medicine Humans Progenitor cell Induced pluripotent stem cell Brain function General Immunology and Microbiology Chemistry General Neuroscience Endothelial Cells Biological Transport Cell Differentiation Cell and molecular biology 030104 developmental biology medicine.anatomical_structure nervous system Blood-Brain Barrier Astrocytes Paracellular transport cardiovascular system Neuroscience 030217 neurology & neurosurgery |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/60925 |
| Popis: | The blood brain barrier (BBB) is formed by neurovascular units (NVUs) that shield the central nervous system (CNS) from a range of factors found in the blood that can disrupt delicate brain function. As such, the BBB is a major obstacle to the delivery of therapeutics to the CNS. Accumulating evidence suggests that the BBB plays a key role in the onset and progression of neurological diseases. Thus, there is a tremendous need for a BBB model that can predict penetration of CNS-targeted drugs as well as elucidate the BBB's role in health and disease. We have recently combined organ-on-chip and induced pluripotent stem cell (iPSC) technologies to generate a BBB chip fully personalized to humans. This novel platform displays cellular, molecular, and physiological properties that are suitable for the prediction of drug and molecule transport across the human BBB. Furthermore, using patient-specific BBB chips, we have generated models of neurological disease and demonstrated the potential for personalized predictive medicine applications. Provided here is a detailed protocol demonstrating how to generate iPSC-derived BBB chips, beginning with differentiation of iPSC-derived brain microvascular endothelial cells (iBMECs) and resulting in mixed neural cultures containing neural progenitors, differentiated neurons, and astrocytes. Also described is a procedure for seeding cells into the organ chip and culturing of the BBB chips under controlled laminar flow. Lastly, detailed descriptions of BBB chip analyses are provided, including paracellular permeability assays for assessing drug and molecule permeability as well as immunocytochemical methods for determining the composition of cell types within the chip. |
| Databáze: | OpenAIRE |
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