Evaluation of the effect of compounds on the integrity of the intestinal barrier

Introduction

The effect of compounds on the integrity of the intestinal barrier can be assessed by measuring the change in resistance of a monolayer of Caco-2 cells after addition of the compound.
The Caco-2 cell line (derived from human colorectal carcinoma) is the model used in vitro to characterise the effects of compounds on the intestinal barrier.

Principle

When grown on semi-permeable membranes, Caco-2 cells differentiate to form a highly functionalized epithelial barrier with remarkable morphological and biochemical similarity to the intestinal epithelium.
Caco-2 cells are grown on a membrane placed between two compartments.
The compounds are added to the apical side of the cell monolayer.
The effect of the compounds is assessed by measuring the trans-epithelial electrical resistance of the cell monolayer.

Protocol

Format24-well format
Cell model

Caco-2 (ATCC HTB-37) differentiated

Reference molecules

IFNɣ + TNFα

Treatment 2 concentrations - 3 points
Incubation

72H à 37°C under 5% CO

Analysismeasurement of cell resistance (TEER) over time

Notes :

  • Provide a minimum of 20 µL at 10 mM of test compounds

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