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

STAR Protoc. 2024 Dec 20;5(4):103416. doi: 10.1016/j.xpro.2024.103416. Epub 2024 Nov 1.

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.1.

Keywords: cell culture; health sciences; immunology.

MeSH terms

  • Cell Survival
  • Coculture Techniques* / methods
  • HT29 Cells
  • Humans
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / metabolism
  • Leukocytes, Mononuclear* / cytology
  • Leukocytes, Mononuclear* / immunology
  • Leukocytes, Mononuclear* / metabolism
  • Permeability