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Updated: Sep 23, 2025
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Investigating Intestinal Barrier Breakdown in Living Organoids
Published on: March 26, 2020
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A permeability assay for mouse intestinal organoids.
Stijn Aaron den Daas1,2, Ugo Soffientini1,2, Shilpa Chokshi1,2
1Roger Williams Institute of Hepatology, Foundation for Liver Research, London SE5 9NT, UK.
STAR Protocols
|May 11, 2022
Summary
We developed a new assay to measure intestinal permeability using mouse organoids. This method detects dead cells entering the organoid lumen to assess the impact of various treatments on gut barrier function.
Area of Science:
- Gastroenterology
- Cell Biology
- Toxicology
Background:
- Intestinal permeability is crucial for gut health.
- Assessing barrier function is vital for understanding disease and treatment effects.
- Existing methods may have limitations in organoid systems.
Purpose of the Study:
- To establish a novel assay for quantifying intestinal permeability in mouse intestinal organoids.
- To provide a method adaptable for various species and research applications.
- To evaluate the impact of external factors on the intestinal barrier.
Main Methods:
- Utilized propidium iodide (PI) as a cell viability indicator.
- Developed an assay based on PI's inability to cross intact cell membranes.
- Measured PI-stained dead cells within the organoid lumen.
Main Results:
- The assay successfully detected increased permeability upon disruption of tight junctions or cell death.
- Quantified PI-stained cells within the lumen correlated with barrier compromise.
- Demonstrated the assay's utility in analyzing effects of insults, toxins, and treatments.
Conclusions:
- The described assay offers a robust method for assessing intestinal organoid permeability.
- This technique can be used to study the effects of various agents on gut barrier integrity.
- The assay is adaptable for broader applications in intestinal research.
