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Related Experiment Video

Updated: Jul 7, 2025

Organoid-Derived Epithelial Monolayer: A Clinically Relevant In Vitro Model for Intestinal Barrier Function
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Method for Two-Dimensional Epithelial Monolayer Formation Derived from Mouse Three-Dimensional Small Intestinal

Yuta Takase1, Toshio Takahashi2

  • 1Suntory Foundation for Life Sciences, Bioorganic Research Institute, Kyoto, Japan. takase@sunbor.or.jp.

Methods in Molecular Biology (Clifton, N.J.)
|December 22, 2023
PubMed
Summary

Researchers developed a simple method to create 2D intestinal epithelial monolayers from 3D organoids. This new platform allows for better study of intestinal stem cells and epithelium physiology.

Keywords:
2D epithelial monolayer3D organoidCell differentiationIntestinal stem cellsMouse small intestineSphere-like morphologyTight junctionTransepithelial electrical resistance

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Area of Science:

  • Gastroenterology
  • Cell Biology
  • Tissue Engineering

Background:

  • The intestinal epithelium's high regenerative capacity relies on intestinal stem cells (ISCs) located in crypts.
  • Three-dimensional (3D) organoid cultures allow study of ISC mechanisms but limit access to the apical epithelium.
  • Established two-dimensional (2D) culture methods for mouse small intestine are lacking.

Purpose of the Study:

  • To develop a simple and effective method for generating 2D epithelial monolayers from mouse 3D small intestinal organoids.
  • To establish a novel platform for studying intestinal epithelium physiology and interactions.

Main Methods:

  • Utilized commercially available materials for 2D monolayer formation from 3D mouse small intestinal organoids.
  • Cultured organoids to form confluent 2D epithelial monolayers within 4 days.

Main Results:

  • Successfully established confluent 2D epithelial monolayers from mouse 3D organoids.
  • The 2D monolayers exhibited stable tight junctions and contained both ISCs and differentiated intestinal cells.
  • Physiologically relevant transepithelial electrical resistance values were observed.

Conclusions:

  • The developed method provides a simple and accessible platform for creating 2D mouse intestinal epithelial models.
  • This 2D culture system facilitates research into intestinal epithelium physiology, host-microbe interactions, and villus formation mechanisms.