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Updated: Dec 14, 2025
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Organoid-Derived Epithelial Monolayer: A Clinically Relevant In Vitro Model for Intestinal Barrier Function
Published on: July 29, 2021
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Primary Intestinal Epithelial Organoid Culture
Tomohiro Mizutani1,2, Hans Clevers3,4,5,6
1Hubrecht Institute, KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands.
Methods in Molecular Biology (Clifton, N.J.)
|July 25, 2020
Summary
Researchers developed a novel method to culture intestinal stem cells, creating 3D organoids that mimic the gut
Area of Science:
- Gastroenterology
- Stem Cell Biology
- Regenerative Medicine
Background:
- The intestinal epithelium exhibits high regenerative capacity, driven by Lgr5+ stem cells in crypts.
- The crypt-villus architecture provides a model for studying stem cell dynamics.
- Understanding stem cell niche signals is crucial for in vitro culture development.
Purpose of the Study:
- To establish a robust in vitro culture method for intestinal epithelial stem cells.
- To create self-renewing 3D intestinal organoids that recapitulate in vivo architecture.
- To provide a versatile model for studying intestinal biology and disease.
Main Methods:
- Isolation of single Lgr5+ intestinal stem cells (ISCs) or whole crypts.
- Culture of ISCs and crypts in specific conditions to promote self-organization.
- Characterization of 3D organoid structures for recapitulation of crypt-villus organization.
Main Results:
- Efficient expansion of single ISCs and whole crypts into 3D organoids.
- Organoids mimic the intestinal crypt-villus architecture.
- Organoids are stable in culture, can be passaged weekly, maintained long-term, and cryopreserved.
Conclusions:
- The established culture method enables long-term expansion of functional intestinal organoids.
- These organoids serve as a powerful tool for studying stem cell biology, gene function, and disease mechanisms.
- Intestinal organoids offer a renewable and manipulable model for gastrointestinal research.
