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Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium
JoVE 杂志
免疫与感染
This content is Free Access.
JoVE 杂志 免疫与感染
Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium

Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium

DOI:
10.3791/59357-v

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07:20 min

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April 09, 2019

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, , ,
1Department of Internal Medicine,University of New Mexico Health Science Center, 2Department of Medicine, Division of Gastroenterology & Hepatology,Johns Hopkins University School of Medicine

Chapters

  • 00:04Title
  • 01:22Isolation and Dissociation of Enteroids/Colonoids from 3D Culture
  • 02:39Plating the Enteroid/Colonoid Suspension
  • 04:10Fixation to Preserve and Immunostain Mucus
  • 05:16Results: Establishment and Evaluation of Human Enteroid/Colonoid Monolayers
  • 06:51Conclusion

Summary

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Here, we present a protocol to culture human enteroid or colonoid monolayers that have intact barrier function to study host epithelial-microbiota interactions at the cellular and biochemical level.

Tags

Colonoid MonolayersIntestinal EpitheliumHost-pathogen InteractionMucus BiologyMicrobiomeInflammatory Bowel Disease3D Organoid CultureEx Vivo StudiesHarvesting SolutionCell SuspensionDMEM/F12 MediumCell PlatingExpansion Medium

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