4.4 Article

In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability

Journal

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 140, Pages -

Publisher

JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/57032

Keywords

Biology; Issue 140; Intestinal barrier function; intestinal permeability; transepithelial electrical resistance; FITC-dextran; Caco-2BBe cells; gavage

Funding

  1. National Natural Science Foundation of China [81470804, 31401229, 81200620]
  2. Natural Science Foundation of Jiangsu Province [BK20180838, BK20140319]
  3. Research Innovation Program for College Graduates of Jiangsu Province [KYLX16-0116]
  4. Advanced Research Projects of Soochow University [SDY2015_06]
  5. Crohn's & Colitis Foundation Research Fellowship Award [310801]

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The intestinal barrier defends against pathogenic microorganism and microbial toxin. Its function is regulated by tight junction permeability and epithelial cell integrity, and disruption of the intestinal barrier function contributes to progression of gastrointestinal and systemic disease. Two simple methods are described here to measure the permeability of intestinal epithelium. In vitro, Caco-2BBe cells are plated in tissue culture wells as a monolayer and transepithelial electrical resistance (TER) can be measured by an epithelial (volt/ohm) meter. This method is convincing because of its user-friendly operation and repeatability. In vivo, mice are gavaged with 4 kDa fluorescein isothiocyanate (FITC)-dextran, and the FITC-dextran concentrations are measured in collected serum samples from mice to determine the epithelial permeability. Oral gavage provides an accurate dose, and therefore is the preferred method to measure the intestinal permeability in vivo. Taken together, these two methods can measure the permeability of the intestinal epithelium in vitro and in vivo, and hence be used to study the connection between diseases and barrier function.

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