Comparative Effects of Culture Media and Cytokine Signaling on J774A.1 Macrophage Activation and Functional Plasticity
Location
Sydnor Performance Hall, Schewel Hall
Access Type
Campus Access Only
Presentation Type
Oral presentation
Entry Number
100
Start Date
4-16-2026 2:30 PM
End Date
4-16-2026 2:45 PM
School
School of Medicine and Health Sciences
Abstract
This research project investigates how culture conditions influence the immune behavior of J774A.1 mouse macrophages, a commonly used model for studying inflammation. Macrophages are critical cells of the innate immune system that detect pathogens and regulate immune responses through signaling molecules such as cytokines and nitric oxide. Previous studies using RAW 264.7 macrophages have shown that media composition, particularly nutrient availability, can significantly alter macrophage function. However, similar studies examining how culture conditions influence both inflammatory (M1) and anti-inflammatory (M2) activation states in J774A.1 macrophages remain limited. The goal of this project is to determine how differences in nutrient composition and immune signaling influence macrophage activation and functional responses. To address this question, J774A.1 macrophages will be cultured in two commonly used media types, DMEM and RPMI 1640, including a glucose-supplemented RPMI condition to match DMEM levels. Cells will be stimulated with bacterial lipopolysaccharide to measure pro-inflammatory responses through nitrite production and cytokine release. Cells will be treated with Interleukin-4 (IL-4) to induce M2 phenotype. Initial work showed significant differences in J774A.1 cells grown in high and low glucose media. Production of the cytokine CCL17 will be measured using an ELISA assay to evaluate M2 macrophage activation. This project will help clarify how culture conditions influence macrophage immune behavior and may improve experimental consistency across laboratories. A better understanding of macrophage functional plasticity may also contribute to future research on inflammatory diseases and immune regulation.
Primary Faculty Mentor(s)
David Freier
Primary Faculty Mentor(s) Department
Biology
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Comparative Effects of Culture Media and Cytokine Signaling on J774A.1 Macrophage Activation and Functional Plasticity
Sydnor Performance Hall, Schewel Hall
This research project investigates how culture conditions influence the immune behavior of J774A.1 mouse macrophages, a commonly used model for studying inflammation. Macrophages are critical cells of the innate immune system that detect pathogens and regulate immune responses through signaling molecules such as cytokines and nitric oxide. Previous studies using RAW 264.7 macrophages have shown that media composition, particularly nutrient availability, can significantly alter macrophage function. However, similar studies examining how culture conditions influence both inflammatory (M1) and anti-inflammatory (M2) activation states in J774A.1 macrophages remain limited. The goal of this project is to determine how differences in nutrient composition and immune signaling influence macrophage activation and functional responses. To address this question, J774A.1 macrophages will be cultured in two commonly used media types, DMEM and RPMI 1640, including a glucose-supplemented RPMI condition to match DMEM levels. Cells will be stimulated with bacterial lipopolysaccharide to measure pro-inflammatory responses through nitrite production and cytokine release. Cells will be treated with Interleukin-4 (IL-4) to induce M2 phenotype. Initial work showed significant differences in J774A.1 cells grown in high and low glucose media. Production of the cytokine CCL17 will be measured using an ELISA assay to evaluate M2 macrophage activation. This project will help clarify how culture conditions influence macrophage immune behavior and may improve experimental consistency across laboratories. A better understanding of macrophage functional plasticity may also contribute to future research on inflammatory diseases and immune regulation.