Article1 min read·Published Oct 2015Older

In vivo Fluorescein Isothiocyanate-dextran (FD4) Permeability Assay

Part of Intestinal Organoid Science & Bioengineering 49 items

pubmed.ncbi.nlm.nih.gov

Michael Helmrath

Using pluripotent stem cells, it is now becoming possible to develop tissue models of organ systems within the body. These organs will allow for the study of organ function, physiology, embryology, and even pathologic processes. Recently, our group developed a model of human small intestine developed from human pluripotent stem cells which when transplanted in vivo, produce a mature, cystic intestinal structure that has digestive functions similar to that of native small intestine (Watson et al., 2014). Intestinal permeability is a primordial function of both the epithelium and associated tight junctions to control nutrient intake and prevent the passage of pathogens. One way to study gastrointestinal paracellular permeability is by determining the ability of fluorophores-conjugated macromolecules (i.e., fluorescein isothiocyanate-dextran (FITC-dextran; or FD4) to cross from the lumen and into circulation (Dong et al., 2014). We were able to test the intestinal permeability by injecting FITC-dextran directly into the lumen of the bioengineered intestine and determining the fluorescence within the blood of the murine host at various time points after injection.

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Article · Oct 2015 · 1 min read

In brief

This protocol describes an in vivo assay to measure intestinal barrier function in bioengineered human intestinal tissue derived from pluripotent stem cells and transplanted into mice. By injecting fluorescent FITC-dextran into the intestinal lumen and measuring blood fluorescence over time, researchers can assess epithelial permeability and tight junction integrity in these organoid models.

Written by the GCMD Library team from the article.

Michael Helmrath

Statements in this article

Pluripotent stem cells can be used to develop tissue models of organ systems for studying organ function, physiology, embryology, and pathologic processes.
Established
Human pluripotent stem cells can generate mature, cystic intestinal structures with digestive functions similar to native small intestine when transplanted in vivo.
Research
Intestinal permeability reflects the function of the epithelium and tight junctions to control nutrient intake and prevent pathogen passage.
Established
Gastrointestinal paracellular permeability can be studied by measuring the ability of fluorescein isothiocyanate-dextran (FITC-dextran or FD4) to cross from the lumen into circulation.
Established
Intestinal permeability in bioengineered intestine can be tested by injecting FITC-dextran directly into the lumen and measuring fluorescence in the murine host blood at various time points.
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