Primary cortical cell tri-culture to study effects of amyloid-β on microglia function and neuroinflammatory response
Year: 2024
Type: article
Abstract: BACKGROUND: Microglia play a critical role in neurodegenerative disorders, such as Alzheimer's disease, where alterations in microglial function may result in pathogenic amyloid-β (Aβ) accumulation, chronic neuroinflammation, and deleterious effects on neuronal function. However, studying these complex factors in vivo, where numerous confounding processes exist, is challenging, and until recently, in vitro models have not allowed sustained culture of critical cell types in the same culture. OBJECTIVE: We employed a rat primary tri-culture (neurons, astrocytes, and microglia) model and compared it to co-culture (neurons and astrocytes) and mono-culture (microglia) to study microglial function (i.e., motility and Aβ clearance) and proteomic response to exogenous Aβ. METHODS: The cultures were exposed to fluorescently-labeled Aβ (FITC-Aβ) particles for varying durations. Epifluorescence microscopy images were analyzed to quantify the number of FITC-Aβ particles and assess cytomorphological features. Cytokine profiles from conditioned media were obtained. Live-cell imaging was employed to extract microglia motility parameters. RESULTS: FITC-Aβ particles were more effectively cleared in the tri-culture compared to the co-culture. This was attributed to microglia engulfing FITC-Aβ particles, as confirmed via epifluorescence and confocal microscopy. FITC-Aβ treatment significantly increased microglia size, but had no significant effect on neuronal surface coverage or astrocyte size. Upon FITC-Aβ treatment, there was a significant increase in proinflammatory cytokines in tri-culture, but not in co-culture. Aβ treatment altered microglia motility evident as a swarming-like motion. CONCLUSIONS: The results suggest that neuron-astrocyte-microglia interactions influence microglia function and highlight the utility of the tri-culture model for studies of neuroinflammation, neurodegeneration, and cell-cell communication.
Source: Journal of Alzheimer s Disease
Institution: University of California, Davis
Language: English
Subfield: Neurology
Field: Neuroscience
Domain: Life Sciences
Open Access status: green
Funders: National Science Foundation, National Institutes of Health, University of California, Davis, National Institute on Aging, National Eye Institute, National Institute of Neurological Disorders and Stroke, National Institute of Environmental Health Sciences, National Institute of Biomedical Imaging and Bioengineering, Division of Materials Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1148898704, G1859492571, G2033485602, G266965019, Experimental and Computational Study of Pore Morphology Evolution Mechanisms in Nanoporous Metal Thin Films Under Thermal/Electrical/Mechanical Stress Fields, G2965285717, G5481912773, G6777974675, G8682997308, G9000011351, G9000036978, G9000048651, G9000067536, G9000117709, G9000120125, G9000120814, G9000121558, G9000226244, G9000334993, G9000352264, UC Davis Environmental Health Sciences Core Center
OpenAlex ID: W4404122207