A simplified cell culture model for research on intestinal inflammation

Abstract

Background/aim: The aim of this study was to combine some easy and economical defined methods and constitute a comprehensive cell culture model to use in the bowel diseases characterized by inflammation. Materials and methods: Induction of inflammation was performed using lipopolysaccharide (LPS) or a cytokine mixture (TNF-α: 10 ng/mL; IFN-γ: 100 ng/mL; IL-1β: 1 ng/mL). IEC-6 cells were grown to confluence and serum-starved, wounds were constituted, and progress of cell migration into the wounds was photographed at 0, 2, 4, 6, 8, 10, 12, and 24 h in the presence or absence of an inflammatory environment. Cells were then grown and multiple scratches were performed to replicate the conditions of migration assay. Nitric oxide synthase-2 (iNOS) and cyclooxygenase-2 (COX-2) protein expressions were assessed. Results: Cells covered 88% of the initial wound at 24 h in the control group, 54% in the LPS group, and 35% in the cytokine mixture group. LPS and the cytokine mixture were also found to independently increase iNOS and COX-2 expressions. Conclusion: Our study, being inexpensive and practical, describes a model that integrates some methods to constitute a basic model for bowel diseases characterized by inflammation. It can be integrated as a preliminary experiment for etiopathogenesis and drug research studies.