Cortical Spheroid Model for Studying the Effects of Ischemic Brain Injury

Abstract

Stroke is a devastating neurological disorder and a leading cause of death and long-term disability. Despite many decades of research, there are still very few therapeutic options for patients suffering from stroke or its consequences. This is partially due to the limitations of current research models, including traditional in vitro models which lack the three-dimensional (3D) architecture and cellular make-up of the in vivo brain. 3D spheroids derived from primary postnatal rat cortex provide an in vivo-relevant model containing a similar cellular composition to the native cortex and a cell-synthesized extracellular matrix. These spheroids are cost-effective, highly reproducible, and can be produced in a high-throughput manner, making this model an ideal candidate for screening potential therapeutics. To study the cellular and molecular mechanisms of stroke in this model, spheroids were deprived of glucose, oxygen, or both oxygen and glucose for 24 hours. Both oxygen and oxygen-glucose deprived spheroids demonstrated many of the hallmarks of stroke, including a decrease in metabolism, an increase in neural dysfunction, and an increase in reactive astrocytes. Pretreatment of spheroids with the antioxidant agent N-acetylcysteine (NAC) mitigated the decrease in ATP seen after 24 hours of oxygen-glucose deprivation. Together, these results show the utility of our 3D cortical spheroid model for studying ischemic injury and its potential for screening stroke therapeutics.

Cortical spheroids show strain-dependent cell viability loss and neurite disruption following sustained compression injury

“…The spheroids contained all the key cell types of the brain, including glia; the cells and neurites were interconnected; they produced their own extracellular matrix; and the neurons were electrically active, with mature synapses (20,32). We have recently demonstrated the utility of these cortical spheroids for modeling disease and injury effects, as they can respond to oxygen deprivation and ischemia (33). While the spheroid does not provide the layer-specific organization of the in vivo cortex, it is an efficient and importantly, a high-throughput approach that presents many key in vivo characteristics.…”