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Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium
Published on: April 9, 2019
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Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium
Julie G In1, Jennifer Foulke-Abel2, Elizabeth Clarke1
1Department of Internal Medicine, University of New Mexico Health Science Center.
Journal of Visualized Experiments : Jove
|April 30, 2019
Summary
Human intestinal organoids, or enteroids/colonoids, are advanced ex vivo models. Modified enteroid/colonoid monolayers improve host-pathogen interaction studies by enhancing surface accessibility.
Area of Science:
- Gastroenterology
- Microbiology
- Stem Cell Biology
Background:
- Human 3D enteroid/colonoid cultures are advanced intestinal epithelium models.
- Their structure limits host-pathogen interaction studies.
- Monolayer cultures offer improved accessibility for research.
Purpose of the Study:
- To adapt 3D enteroids/colonoids into accessible monolayers.
- To establish a model for studying host-pathogen interactions.
- To enable investigation of microbiota-epithelial cell interactions.
Main Methods:
- Fragmentation of 3D cultures and seeding onto permeable membranes.
- Monitoring confluency and differentiation using transepithelial electrical resistance (TER).
- Analysis of secreted mucus via immunofluorescence or immunoblotting.
Main Results:
- Successful generation of colonoid monolayers from 3D cultures.
- TER measurements indicated successful differentiation and confluency.
- Secreted mucus from colonoid monolayers was characterized.
Conclusions:
- Enteroid/colonoid monolayers provide a physiologically relevant platform.
- This model enhances studies of bacterial-host epithelial interactions.
- The platform facilitates evaluation of host cell targeting by microbiota.
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