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An ex vivo method for studying mucus formation, properties and thickness in human colonic biopsies and mouse small and large intestinal explants.

Journal article
Authors Jenny K Gustafsson
Anna Ermund
Malin E V Johansson
André Schütte
Gunnar C. Hansson
Henrik Sjövall
Published in American journal of physiology. Gastrointestinal and liver physiology
Volume 302
Issue 4
Pages 430-438
ISSN 1522-1547
Publication year 2012
Published at Institute of Medicine, Department of Internal Medicine and Clinical Nutrition
Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 430-438
Language en
Links dx.doi.org/10.1152/ajpgi.00405.2011
Keywords mucin colon ulcerative colitis
Subject categories Gastroenterology and Hepatology

Abstract

The colon mucus layers minimize the contact between the luminal flora and the epithelial cells and defects in this barrier may lead to colonic inflammation. We now describe an ex vivo method for analysis of mucus properties in human colon and mouse small and large intestine. Materials and methods: Intestinal explants were mounted in horizontal perfusion chambers. The mucus surface was visualized by adding charcoal particles on the apical side and mucus thickness was measured using a micropipette. Mucus thickness, adhesion and growth rate was recorded for 1 h. In mouse and human colon, the ability of the mucus to act as a barrier to beads the size of bacteria was also evaluated. Tissue viability was monitored by transepithelial potential difference. Results: In mouse ileum the mucus could be removed by gentle aspiration, whereas in colon about 40 µm of the mucus remained attached to the epithelial surface. Both mouse and human colon had an inner mucus layer that was not penetrated by the fluorescent beads. Spontaneous mucus growth was observed in human (240 µm/h) and mouse (100 µm/h) colon, but not in mouse ileum. In contrast, stimulation with carbachol induced a higher mucus secretion in ileum than colon (mouse ileum: Δ200 μm, mouse colon: Δ130 µm, human colon: Δ140 μm). In conclusion, while retaining key properties from the mucus system in vivo, this set up also allows for studies of the highly dynamic mucus system under well controlled conditions.

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