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Updated: Nov 15, 2025
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Co-Culture of Murine Small Intestine Epithelial Organoids with Innate Lymphoid Cells
Published on: March 23, 2022
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In vitro Self-organized Mouse Small Intestinal Epithelial Monolayer Protocol.
Gizem Altay1, Eduard Batlle2,3,4, Vanesa Fernández-Majada1
1Biomimetic Systems for Cell Engineering Laboratory, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.
Bio-Protocol
|March 3, 2021
Summary
Researchers developed a simple method for creating intestinal epithelial monolayers that mimic in vivo conditions. This model supports stem cell function and barrier properties for various functional studies.
Area of Science:
- Gastroenterology and Hepatology
- Cell Biology
- Biomedical Engineering
Background:
- Organoid models have limitations due to their closed geometry, hindering physiological recapitulation.
- Existing culture models often lack self-renewing capacities despite physiological cell composition.
Purpose of the Study:
- To develop a novel method for generating intestinal epithelial monolayers with in vivo-like architecture and function.
- To establish a model suitable for functional assays requiring access to both luminal and basolateral compartments.
Main Methods:
- Isolation of mouse small intestine epithelial cells.
- Culture of cells to form self-renewing monolayers with crypt-like and villus-like domains.
- Adaptation of the model to a tissue culture format for functional studies.
Main Results:
- Successfully generated intestinal epithelial monolayers with proliferative crypt-like domains and differentiated villus-like regions.
- Demonstrated close-to-physiological barrier properties of the in vitro monolayers.
- Confirmed presence of stem cells and appropriate cell distribution mirroring in vivo conditions.
Conclusions:
- The developed protocol yields physiologically relevant intestinal epithelial monolayers.
- This model overcomes organoid limitations and is suitable for drug absorption, intracellular trafficking, and microbiome-epithelium interaction studies.
