A simplified cell culture model for research on intestinal inflammation

WOS: 000431261100035PubMed ID: 29714467Background/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-alpha: 10 ng/mL; IFN-gamma: 100 ng/mL; IL-1 beta: 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.Research Foundation of Ege University, Izmir, TurkeyEge University [2012-TIP-053]This study was supported by the Research Foundation of Ege University (Project No: 2012-TIP-053), Izmir, Turkey. The authors thanks Prof Dr Mehmet Orman (Ege University, Faculty of Medicine, Department of Biostatistics) for his contribution in statistical analysis of the study data. Some of the data given in this article were presented as a poster presentation at the 22nd National Congress of Pharmacology, Antalya, Turkey