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Name: Personal
Name: Personal
Name: Personal
Name: Personal
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Note: thesis: Thesis (Sc. M.)--Brown University, 2017
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Abstract: Central nervous system disease pathologies are poorly understood in part due to the inability of animal and in vitro models to recapitulate their complexity. It is therefore crucial to develop three-dimensional in vitro models of the central nervous system in which cells demonstrate more in-vivo-like gene and protein expression relative to traditional 2D culture. We previously described a primary cortical spheroid model that enables the in vitro study of in vivo-relevant characteristics, such as cell density, cell composition complexity, neuronal electrophysiology, tissue stiffness, and dimensionality. Importantly, cortical endothelial cells spontaneously assemble into capillary-like network structures within cortical spheroids, allowing for the study of the neurovasculature. Immunohistochemistry revealed that capillary-like networks are surrounded by basement membrane proteins and interact with relevant neural cell types. The networks are dynamic and change structure over the course of their lifetime, responding to the introduction of other cell types. This model provides a 3D scaffold-free environment to study interactions between the complex cells of the neurovascular unit in an in vitro setting. Additionally, the dysfunction of the neurovasculature in central nervous system diseases makes this model applicable to investigating the vasculature in both healthy and diseased states. A disease state of interest, ischemic stroke, is a major cause of morbidity and mortality worldwide and occurs when there is a localized reduction in regional blood flow in the brain resulting in an undersupply of oxygen and nutrients. We have begun to develop and characterize a three-dimensional in vitro model of ischemic stroke using primary cortical spheroids. The spheroids were deprived of oxygen and glucose for 24 hours using glucose-free media and anaerobic culturing conditions, and then allowed to recover in normoxic conditions to model reperfusion. The extent of injury was characterized using the LDH cell viability assay optimized to our cortical spheroids and immunohistochemistry. Age and size of the cortical spheroids impacts the resulting cell viability after ischemic injury. Additionally, ischemic injury affects various neuronal cell types present in the spheroids. The capillary-like networks are dynamic and responded to ischemic injury by losing their tubular structures and forming rings in the center of the spheroids. Further development of this injury model provides the potential for a relevant and tailorable in vitro platform for ischemic injury.
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Subject (fast) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/00851293")
Subject (fast) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01036575")
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Identifier: DOI: 10.7301/Z0FT8JHR
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Access Condition: restriction on access: Collection is open for research.