In-vitro model for gut epithelial barrier (GLP-like / GLP)

We provide a cell-based model of the human gut epithelial barrier to analyze the impact of test substances and/or infectious agents on gut barrier integrity and function. Caco-2 cells (human epithelial cell line) and THP-1 cells (human monocytic cell line) are co-cultured in a transwell culture system that models the epithelial barrier (Caco-2 cells) and the underlying lamina propria with resident macrophages (THP-1 cells). After addition of test substances and/or infectious agents to the upper chamber of the transwell system ("gut lumen"), epithelial barrier integrity and function as well as transmigration of pathogens and the resulting immune response by epithelial cells and macrophages is analyzed using various approaches.

 

In-vitro model for gut epithelial barrier (GLP-like / GLP)

 

Fig.: Experimental setup and analytical endpoints of the in-vitro model for gut epithelial barrier and lamina propria. The expression of tissue markers (e.g. Occludin) in the Caco-2 cell monolayer is analyzed by immunofluorescence (IF) followed by quantification. An ultra-structural analysis can be obtained by transmission electron microscopy (TEM). The small arrow indicates a tight junction.

Species

Human

Fields of application

  • Preclinical drug development
  • Intestinal infection studies
  • Toxicity studies
  • Efficacy and safety studies for drug approval (GLP-like and GLP)

Endpoints/Outcome parameter

  • Barrier integrity / permeability
  • Cell viability / cytotoxicity
  • Pathogen invasiveness / migration
  • Profile of cytokines / chemokines
  • Changes in tissue marker expression (e.g. tight junction proteins)
  • Ultra-structural analysis of epithelial barrier

Readout parameter

  • TEER (Transepithelial electrical resistance)
  • FITC-Dextran test
  • Cell viability / cytotoxicity assays
  • CFU assays
  • ELISA / Multiplex analysis
  • Histology
  • Immunohistochemistry / Immunofluorescence
  • Transmission electron microscopy

Quality management and validation

  • Internal quality management (certified GLP test facility)
  • Use of reference compounds
  • Randomisation
  • Blinded induction, data acquisition and analysis
  • Biometric expertise