Developing a Three-Dimensional Brain Microtissue Model of Ischemic-Reperfusion Injury

McLaughlin, Rachel M.

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Overview

Year:: 2023
Contributor:: McLaughlin, Rachel M (creator); Hoffman-Kim, Diane (Advisor); Lipscombe, Diane (Reader); Webb, Ashley (Reader); Thorne, Robert (Reader); Brown University. Department of Neuroscience (sponsor)
Genre:: theses
Subject:: Image analysis; Brain; Stroke; 3D Cell Culture; ischemic-reperfusion; Central nervous system
Extent:: 16, 208 p.

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Abstract:: Stroke is a devastating medical emergency caused by a lack of blood flow to a particular area of the brain. Even short interruptions in blood flow can lead to devastating consequences. Despite many decades of research, there are still very few therapeutic options for patients suffering from ischemic stroke. In this thesis, we aimed to develop a model of ischemic-reperfusion injury using our previously established 3D microtissues derived from primary postnatal rat cortex and develop a method for analyzing complex biological images. The cortical microtissues provide an in vivo-relevant model containing a similar cellular composition to the native cortex and a cell-synthesized extracellular matrix. This model is cost-effective, highly reproducible, and can be produced in a high-throughput manner, making it an ideal candidate for screening potential therapeutics. To study the cellular and functional mechanisms of ischemic and reperfusion injury in this model, spheroids were deprived of glucose, oxygen, or both oxygen and glucose for 24 hours and reintroduced to normal culture conditions for an additional 24 hours. Both oxygen and oxygen-glucose deprived spheroids demonstrated many of the hallmarks of ischemic brain injury, including a decrease in metabolism, an increase in neural dysfunction, breakdown in the neurovascular unit, and an increase in reactive astrocytes. After reperfusion, there was an increase in reactive oxygen species production and increase in cell death. Neural cytoskeletal structure and capillary-like networks continue to be disrupted, but there are some signs of astrocyte and calcium dynamic recovery. Pretreatment of spheroids with the antioxidant agent N-acetylcysteine (NAC) mitigated the decrease in ATP after oxygen-glucose deprivation, was partially neuroprotective, and enhanced the expression of laminin. Together, these results show the utility of our 3D cortical microtissue model for studying ischemic-reperfusion injury and its potential for screening stroke therapeutics.
Notes:: Thesis (Ph. D.)--Brown University, 2023

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Citation

McLaughlin, Rachel M., "Developing a Three-Dimensional Brain Microtissue Model of Ischemic-Reperfusion Injury" (2023). Neuroscience Theses and Dissertations. Brown Digital Repository. Brown University Library. https://repository.library.brown.edu/studio/item/bdr:eb4zwk76/

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Neuroscience Theses and Dissertations

Theses and Dissertations for the Neuroscience department.
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