Related Experiment Video
Updated: Apr 1, 2026
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Culture Methods to Study Apical-Specific Interactions using Intestinal Organoid Models
Published on: March 23, 2021
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A Two-Dimensional Organoid Culture on Invasin or Basement Membrane Extract to Study Host-Bacteria Interactions
Theodore Grenier1, Katarína Balážová1, Joost J A P M Wijnakker1
1Hubrecht Institute for Developmental Biology and Stem Cell Research: Hubrecht Institute; Oncode Institute; Royal Netherlands Academy of Arts and Science (KNAW); University Medical Center Utrecht.
Journal of Visualized Experiments : Jove
|March 30, 2026
Summary
Tissue stem cell (TSC)-derived organoids offer a powerful in vitro model for studying human physiology and host-microbe interactions. This adaptable technology enables detailed investigation of bacterial infections in human cells.
Area of Science:
- Biotechnology
- Microbiology
- Human Physiology
Background:
- Organoid technology offers an advanced in vitro platform for studying human health and disease.
- Tissue stem cell (TSC)-derived organoids provide advantages over traditional cell lines, including indefinite expansion, genomic stability, and CRISPR compatibility.
- Organoids contain native cell types, making them ideal for studying interactions between human cells and microbes.
Purpose of the Study:
- To establish and validate a versatile organoid model for investigating host-microbe interactions, specifically bacterial infections.
- To demonstrate the utility of intestinal organoids for studying pathogenic bacteria adherence and invasion.
- To provide a protocol adaptable for studying various organs and bacterial species.
Main Methods:
- Generating two-dimensional polarized intestinal organoid monolayers on cell culture inserts.
- Utilizing Basement Membrane Extract (BME) or invasin for coating.
- Differentiating organoids into major intestinal epithelial cell types for infection studies with adherent-invasive E. coli.
- Analyzing infection by monitoring gene expression (RT-qPCR), visualizing interactions (confocal microscopy), and quantifying barrier breaching.
Main Results:
- Intestinal organoids effectively model human epithelial cell interactions with pathogenic bacteria.
- Key infection parameters such as gene expression, bacterial-epithelial interactions, and barrier integrity can be quantified.
- The protocol demonstrates successful infection studies with adherent-invasive E. coli.
Conclusions:
- Organoid technology provides a robust and adaptable platform for studying host-microbe interactions in a human physiological context.
- This model system facilitates research into bacterial pathogenesis and host responses.
- The protocol's adaptability supports broader applications in organoid-based research for infectious diseases.
