The Role of Chrysin in the Modulation of the Immunogenicity of H9 derived Human Neural Progenitor Cells, Neurons and Astrocytes in the presence of LPS
Cardiff Metropolitan University
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Background Inflammation has been known to be a significant contributing factor in the pathophysiology of neurodegenerative diseases for several years. A known causative agent of inflammation is lipopolysaccharide (LPS) as these molecules have been implicated in the induction of the IκB-NF-κβ pathway and the production of inflammatory cytokines such as tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6), which leads to the subsequent activation of the JAK/STAT3 pathway. In recent years neutraceuticals, in particular chrysin, have been shown to modulate the cellular responses to inflammatory stimuli such as LPS in several cell types. The aim of this research is to investigate further into whether the anti-inflammatory properties of chrysin can be mimicked in neural cells and whether there is a difference in response to the supplement between neural progenitor cells, neurons and astrocytes. Methodology NSCs, neurons and astrocytes were grown up into six 12 well plates having been differentiated from H9 derived hESCs. One of each plate of cells went under a Propidium iodide viability assay using flow cytometry for three different concentrations of LPS to determine the optimum concentration to use in subsequent analyses. Each cell type was then treated with either 100ng/ml of LPS, 25μMol of chrysin and a combination of both LPS and chrysin. An Enzyme Linked Immunosorbent Assay (ELISA) was performed to ascertain the concentration of interleukin-6 (IL-6) produced by each cell type under each condition. Results Data obtained from the viability assay showed no statistical evidence of a change in viability across the three concentrations of LPS within each cell type. Therefore the maximum dose tested was used for subsequent analyses. The data obtained from the ELISA analysis showed that there was no statistical difference in the production of IL-6 5 within each cell type, suggesting that chrysin did not alter the immunogenicity of the cells in question. Conclusion In conclusion the results of this research show that the anti-inflammatory properties of the flavonoid chrysin seen in other cell types and animal models could not be mimicked in H9 derived human neural progenitor cells themselves and in neurons and astrocytes derived from them. In addition to this there was no statistical difference noted in response to chrysin within the cell types at the concentrations administered to the neural progenitor cell, neuronal and astrocyte cultures.
BSc Biomedical Science (Hons)
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