A novel tri‐culture model for neuroinflammation

• Published in: Journal of neurochemistry 2021-01, Vol.156 (2), p.249-261
• Main Authors: Zheng, Yan‐Fang, Zhou, Xian, Chang, Dennis, Bhuyan, Deep Jyoti, Zhang, Jie Ping, Yu, Wen‐Zhen, Jiang, Xia‐Sen, Seto, Sai Wang, Yeon, Seung Yeon, Li, Jia, Li, Chun Guang
• Format: Article
• Language: English
• Subjects: Alzheimer's disease; Animals; Biotechnology; Caspase; Cell culture; Cell Culture Techniques; Coculture Techniques; Crosstalk; Cytokines; destructed endothelial barrier; Disease Models, Animal; Endothelial Cells; Inflammation; Interleukins; Lipopolysaccharides; Mental disorders; Mice; Microenvironments; Microglia; microglia activation; Microglial cells; Microvasculature; Movement disorders; Neuroblastoma; Neurodegenerative diseases; neuroinflammation; neuron injury; Neurons; Nitric oxide; Parkinson's disease; Pathogenesis; Permeability; Phosphorylation; Schizophrenia; Signal transduction; Tau protein; Toll-like receptors; tri‐culture; Tumor necrosis factor; Tumors
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/jnc.15171

Neuroinflammation is believed to play a primary role in the pathogenesis of most neurodegenerative diseases including Alzheimer's disease, Parkinson's disease and schizophrenia. Currently, suitable in vitro neuroinflammation models for studying cellular interactions and inflammatory mechanisms at the neurovascular unit are still scarce. In this study, we established an experimentally flexible tri‐culture neuroinflammation model combining murine microglial cells (N11), mouse neuroblastoma Nuro2A cell lines and brain microvascular endothelial MVEC(B3) cells in a transwell co‐culture system stimulated with lipopolysaccharides. Neuroinflammation was induced in this tri‐culture model as manifested by activated N11 cells via toll‐like receptor 4, resulting in increased release of proinflammatory mediators (nitric oxide, interleukin‐6 and tumour necrosis factor‐α) through the activation of nuclear factor‐κB signalling pathway. The released inflammatory cytokines from N11 in turn, damaged the tight junction in microvascular endothelial MVEC(B3) cells, increased permeability of endothelial barrier, and induced tau phosphorylation and up‐regulated caspase‐3 expression in mouse neuroblastoma Nuro2A cell lines, leading to neuroinflammation injury. In summary, this tri‐culture inflammation model mimics the microenvironment, the cellular crosstalk and the molecular events that take place during neuroinflammation. It provides a robust in vitro model for studying neuroinflammation mechanisms and screening for potential therapeutics to treat various neurodegenerative diseases. We have established a novel neuroinflammation tri‐culture model that combines N11 microglia, N2A neurons and brain endothelial MVEC (B3) cells in a transwell tri‐culture system under the stimulation of lipopolysaccharides. By binding to the toll‐like receptor 4, lipopolysaccharides provoked IKK‐NFκB pathway which triggered the release of pro‐inflammatory mediators, and that diminished the endothelial barrier and induced tau phosphorylation and caspase‐3 expressions in neurons. We believe this experimentally flexible tri‐culture model mimicking the microvascular unit in vivo and could be extensively used for drug screening and discovery for potential therapeutic compounds against neuroinflammation induced neurodegenerative diseases.