Protocol for the preclinical evaluation of gut barrier function and immune interaction in an HT-29/PBMC co-culture model.

Anne Abot, Claude Knauf, Patrice D Cani, Nicolas Pomié, Gwendoline Astre, Laurianne Créchet, Ophélie Pereira

Journal: STAR protocols 2024;5(4):103416

PMID: 39487982

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Abstract

Here, we present a protocol for the pre-clinical evaluation of gut barrier function and immune interaction in an HT-29/PBMC co-culture model. We describe steps for culturing HT-29 cells and peripheral blood mononuclear cells (PBMCs), assembling the co-culture model, and testing cell viability. We then detail procedures for performing efficacy tests through stress challenges and barrier permeability assays. For complete details on the use and execution of this protocol, please refer to Abot et al..

Address: Enterosys SAS, Labège, France.; NeuroMicrobiota, International Research Program (IRP) INSERM/UCLouvain, France/Belgium; UCLouvain, Université Catholique de Louvain, Louvain Drug Research Institute (LDRI), Metabolism and Nutrition Research Group (MNUT), Brussels, Belgium; Walloon Excellence in Life Sciences and BIOtechnology (WELBIO), WELBIO Department, WEL Research Institute, Wavre, Belgium; UCLouvain, Université Catholique de Louvain, Institute of Experimental and Clinical Research (IREC), 1200 Brussels, Belgium.; NeuroMicrobiota, International Research Program (IRP) INSERM/UCLouvain, France/Belgium; INSERM U1220, Institut de Recherche en Santé Digestive (IRSD), Université Paul Sabatier, Toulouse III, CHU Purpan, Toulouse, France. Electronic address: [email protected].; Enterosys SAS, Labège, France. Electronic address: [email protected].

DOI: 10.1016/j.xpro.2024.103416

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MeSH Terms: Humans, Coculture Techniques, Leukocytes, Mononuclear, HT29 Cells, Intestinal Mucosa, Cell Survival, Permeability