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Updated: Sep 6, 2025
Testing Epithelial Permeability in Fetal Tissue-Derived Enteroids
Published on: June 16, 2022
Testing Epithelial Permeability in Fetal Tissue-Derived Enteroids
Amelia Llerena1, Shaheda Urmi2, Jahanshah Amin3
1Morsani College of Medicine, University of South Florida.
Insights
Human fetal enteroids model preterm infant intestinal injury. Lipopolysaccharide exposure increased enteroid permeability, supporting dysbiosis as a risk factor.
Area of Science:
- Gastroenterology
- Developmental Biology
- Microbiology
Background:
- Human fetal tissue-derived enteroids offer a novel in vitro model for studying intestinal injuries in preterm infants.
- Intestinal injuries in preterm infants lead to mucosal inflammation and increased permeability, which are difficult to assess directly in patients.
- Enteroids mimic intestinal structure and function, enabling investigation of epithelial permeability and tight junction protein regulation.
Purpose of the Study:
- To establish and characterize human fetal enteroids as a model for intestinal injury.
- To investigate the role of lipopolysaccharide (LPS) in inducing epithelial permeability in enteroids.
- To explore the contribution of gram-negative bacterial dysbiosis to intestinal injury mechanisms in preterm infants.
Main Methods:
- Establishing enteroids from human fetal intestinal tissue.
- Characterizing tight junction proteins using immunofluorescent imaging.
- Quantifying paracellular permeability by microinjecting fluorescein-labeled dextran into enteroid lumens and measuring leakage.
Main Results:
- Apical exposure to lipopolysaccharide (LPS) induced epithelial permeability in enteroids in a concentration-dependent manner.
- Enteroid tight junction proteins were characterized, providing a basis for assessing permeability changes.
- The model successfully demonstrated LPS-induced permeability, mimicking effects of gram-negative bacterial dysbiosis.
Conclusions:
- Human fetal enteroids serve as a viable in vitro model for studying intestinal injuries in preterm infants.
- Lipopolysaccharide exposure increases enteroid epithelial permeability, supporting its role in intestinal injury.
- These findings suggest that gram-negative dominant dysbiosis is a significant contributor to intestinal injury mechanisms in preterm infants.
Abstract:
Human fetal tissue-derived enteroids are emerging as a promising in vitro model to study intestinal injuries in preterm infants. Enteroids exhibit polarity, consisting of a lumen with an apical border, tight junctions, and a basolateral outer layer exposed to growth media. The consequences of intestinal injuries include mucosal inflammation and increased permeability. Testing intestinal permeability in vulnerable preterm human subjects is often not feasible. Thus, an in vitro fetal tissue-derived intestinal model is needed to study intestinal injuries in preterm infants. Enteroids can be used to test changes in epithelial permeability regulated by tight junction proteins. In enteroids, intestinal stem cells differentiate into all epithelial cell types and form a three-dimensional structure on a basement membrane matrix secreted by mouse sarcoma cells. In this article, we describe the methods used for establishing enteroids from fetal intestinal tissue, characterizing the enteroid tight junction proteins with immunofluorescent imaging, and testing epithelial permeability. As gram-negative dominant bacterial dysbiosis is a known risk factor for intestinal injury, we used lipopolysaccharide (LPS), an endotoxin produced by gram-negative bacteria, to induce permeability in the enteroids. Fluorescein-labeled dextran was microinjected into the enteroid lumen, and serial dextran concentrations leaked into the culture media were measured to quantify the changes in paracellular permeability. The experiment showed that apical exposure to LPS induces epithelial permeability in a concentration-dependent manner. These findings support the hypothesis that gram-negative dominant dysbiosis contributes to the mechanism of intestinal injury in preterm infants.
