Inflammatory Cell Models (Coculture)
CellMade Advanced Cell Models models are coculture systems that allow cross-talk between two different cell types without any contact between them. These cell culture systems provide a reliable way to identify the earliest biochemical lipid biomarkers of TNF-α activation in real time, right from the very first initiation of inflammatory response. As a result, CellMade Advanced Cell Models models offer the opportunity to identify early biomarkers of inflammation which may be useful to predicting inflammatory response way ahead of cell phenotypic changes or clinical symptoms. The biomarkers discovered using this approach have the added advantage of possessing future feasibility in the clinical setting, since lipids can be readily measured from peripheral samples such as blood.
Inflammation is involved in a variety of diseases such as Obesity, NAFLD-NASH, Diabetes type II, Cardiovascular Disease and Chronic Obstructive Pulmonary Disease. In order to discover potential treatments for these diseases, further research has to be conducted in suitable in vitro models for inflammation, mimicking human pathophysiology. One potential model is the coculture, where several cell types are cultured together to generate cross-talk in order to better mimic the in vivo situation.
A coculture model employing primary human aortic endothelial cells (HAoEC) and primary human peripheral blood mononuclear cells (hPBMC) in order to mimic vascular inflammation was developed. We here describe functionality of such advanced cell model via various experiments where lipopolysaccharide (LPS) stimulated PBMC were co-cultured with HAoEC, in order to generate an inflammatory response in the HAoEC layer. This model is in use to screen for novel, inflammation specific, lipid biomarkers. It was hypothesized that LPS-induced inflammatory reaction in PBMC will result in a secondary inflammatory response in HAoEC, leading to up-regulation of eicosanoid production and changes in cellular lipid profiles. Using Cytometry and LC-MS, obtained results demonstrated that LPS-activated PBMC generate a secondary inflammatory response in HAoEC characterized by changes in (ceramide) lipid profiles. These results demonstrate biological functionality of the developed coculture model.
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