Multiscale Invasion Assay for Probing Macrophage Response to Gram-Negative Bacteria
Year: 2022
Type: article
Abstract: The immune system is a complex network of various cellular components that must differentiate between pathogenic bacteria and the commensal bacteria of the human microbiome, where misrecognition is linked to inflammatory disorders. Fragments of bacterial cell wall peptidoglycan bind to pattern recognition receptors within macrophages, leading to immune activation. To study this complex process, an approach for three-dimensional (3D) culture of human macrophages and their invasion with relevant bacteria in a well-defined hydrogel-based synthetic matrix inspired by the gut was established. Workflows were developed for monocyte encapsulation and differentiation into macrophages in 3D culture with high viability. Bacteria invaded into macrophages permitted in situ peptidoglycan labeling. Macrophages exhibited biologically-relevant cytokine release in response to bacteria. This multi-dimensional bacteria-macrophage co-culture system will prove useful in future studies to observe bacterial fragment production and localization in the cell at the carbohydrate level for insights into how our immune system properly senses bacteria. TOC Figure
Source: Frontiers in Chemistry
Institutions: University of Delaware, Delaware Biotechnology Institute
Language: English
Subfield: Organic Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $2,950
Funders: National Science Foundation, University of Delaware, Delaware IDeA Network of Biomedical Research Excellence, National Institutes of Health, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of General Medical Sciences, Division of Materials Research, Common Fund
Awards: G1177608224, G1200448030, G1705962760, G1883723449, G2607737512, Metabolic carbohydrate cell wall probes for bacterial structure and immune recognition studies, G2902092823, Discovery of Chemical Probes and Therapeutic Leads, Phase II, G3115553707, G3350574766, G366302655, Unraveling fibrosis with dynamic microenvironments and molecular tools, G4444685404, G4523879168, G4973587063, G545157245, G5575959162, G5641144139, Chemistry-Biology Interface (CBI) NIH T32 Training Grant, G6602124872, G6718912342, G6772302223, University of Delaware MRSEC - Center for Hybrid, Active, and Responsive Materials (CHARM), Molecular Design of Advanced Biomaterials, G9000004818, G9000125231, G9000217648, Delaware INBRE, Chemistry-Biology Interface Predoctoral Training Grant 2024-2029
OpenAlex ID: W4213166694