THE INVESTIGATION OF CYTOKINE SECRETION IN THP-1 DERIVED MICROGLIAL CELLS
Cardiff Metropolitan University
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TNF-α and IL-1β are proinflammatory cytokines, secreted by immune cells all over the human body. TNF-α is required for wound healing and pathogen removal and is responsible for necrosis and cell apoptosis. IL-1β is a prototypic proinflammatory cytokine involved in cell proliferation, differentiation and apoptosis. The aim of this investigation was to determine whether subjecting THP-1 derived microglial cells to stimulus from LPS would alter the concentration of secreted cytokines. THP-1 cells were cultured and, using a neural supplement, differentiated to produce a microglial-like state cell model. Cells were then treated with LPS and incubated for a period of 6 hours. Supernatants were collected and tested using DuoSet ELISAs to determine levels of TNF-α, IL-1β and IL-6 secreted. The cells failed to produce any readable data for IL-6 and statistical analysis indicated that LPS did not have a significant effect on the secretion of neither TNF-α nor IL-1β. This study demonstrated that the microglial cells were able to secrete significant amounts of TNF-α and IL-1β but not in response to LPS and were also unable to secrete measurable amounts of IL-6, particularly over the 6- hour time period. This study suggests that these cells may be acting as an interesting model of microglial cells. This is a slightly promising result as there is a need for good human microglial cell models. Further studies are required to investigate gene expression through methods such as Real-time PCR; to elucidate the mechanisms involved in the cell model and also how these cells are capable of responding to other inflammatory stimuli (gBSA, CML, CLA, IFN-ɣ).
BSc Biomedical Sciences
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