The Ussing chamber system for measuring intestinal permeability in health and disease.

Autor: Thomson A; Institute of Infection and Immunity, School of Medicine, Cardiff, UK. ThomsonAA2@cardiff.ac.uk.; Present address: Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, UK. ThomsonAA2@cardiff.ac.uk., Smart K; Institute of Infection and Immunity, School of Medicine, Cardiff, UK., Somerville MS; Institute of Infection and Immunity, School of Medicine, Cardiff, UK., Lauder SN; Institute of Infection and Immunity, School of Medicine, Cardiff, UK., Appanna G; Institute of Infection and Immunity, School of Medicine, Cardiff, UK.; Department of Gastroenterology and Hepatology, University Hospital Wales, Cardiff, UK., Horwood J; Department of Surgery, University Hospital Wales, Cardiff, UK., Sunder Raj L; Department of Gastroenterology and Hepatology, University Hospital Wales, Cardiff, UK., Srivastava B; Department of Gastroenterology and Hepatology, University Hospital Wales, Cardiff, UK., Durai D; Department of Gastroenterology and Hepatology, University Hospital Wales, Cardiff, UK., Scurr MJ; Institute of Infection and Immunity, School of Medicine, Cardiff, UK., Keita ÅV; Division of Surgery, Orthopedics & Oncology, Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden., Gallimore AM; Institute of Infection and Immunity, School of Medicine, Cardiff, UK., Godkin A; Institute of Infection and Immunity, School of Medicine, Cardiff, UK.; Department of Gastroenterology and Hepatology, University Hospital Wales, Cardiff, UK.
Jazyk: angličtina
Zdroj: BMC gastroenterology [BMC Gastroenterol] 2019 Jun 20; Vol. 19 (1), pp. 98. Date of Electronic Publication: 2019 Jun 20.
DOI: 10.1186/s12876-019-1002-4
Abstrakt: Background: The relationship between intestinal epithelial integrity and the development of intestinal disease is of increasing interest. A reduction in mucosal integrity has been associated with ulcerative colitis, Crohn's disease and potentially could have links with colorectal cancer development. The Ussing chamber system can be utilised as a valuable tool for measuring gut integrity. Here we describe step-by-step methodology required to measure intestinal permeability of both mouse and human colonic tissue samples ex vivo, using the latest equipment and software. This system can be modified to accommodate other tissues.
Methods: An Ussing chamber was constructed and adapted to support both mouse and human tissue to measure intestinal permeability, using paracellular flux and electrical measurements. Two mouse models of intestinal inflammation (dextran sodium sulphate treatment and T regulatory cell depletion using C57BL/6-FoxP3 DTR mice) were used to validate the system along with human colonic biopsy samples.
Results: Distinct regional differences in permeability were consistently identified within mouse and healthy human colon. In particular, mice showed increased permeability in the mid colonic region. In humans the left colon is more permeable than the right. Furthermore, inflammatory conditions induced chemically or due to autoimmunity reduced intestinal integrity, validating the use of the system.
Conclusions: The Ussing chamber has been used for many years to measure barrier function. However, a clear and informative methods paper describing the setup of modern equipment and step-by-step procedure to measure mouse and human intestinal permeability isn't available. The Ussing chamber system methodology we describe provides such detail to guide investigation of gut integrity.
Databáze: MEDLINE
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