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A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction
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A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction
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Journal Article
A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction
2024
Overview
Mechanistic investigation of host-microbe interactions in the human gut are hindered by difficulty of co-culturing microbes with intestinal epithelial cells. On one hand the gut bacteria are a mix of facultative, aerotolerant or obligate anaerobes, while the intestinal epithelium requires oxygen for growth and function. Thus, a coculture system that can recreate these contrasting oxygen requirements is critical step towards our understanding microbial-host interactions in the human gut. Here, we demonstrate
Intestinal Organoid Physoxic Coculture
(IOPC) system, a simple and cost-effective method for coculturing anaerobic intestinal bacteria with human intestinal organoids (HIOs). Using commensal anaerobes with varying degrees of oxygen tolerance, such as nano-aerobe
Bacteroides thetaiotaomicron
and strict anaerobe
Blautia
sp., we demonstrate that IOPC can successfully support 24–48 hours HIO-microbe coculture. The IOPC recapitulates the contrasting oxygen conditions across the intestinal epithelium seen
in viv
o. The IOPC cultured HIOs showed increased barrier integrity, and induced expression of immunomodulatory genes. A transcriptomic analysis suggests that HIOs from different donors show differences in the magnitude of their response to coculture with anaerobic bacteria. Thus, the IOPC system provides a robust coculture setup for investigating host-microbe interactions in complex, patient-derived intestinal tissues, that can facilitate the study of mechanisms underlying the role of the microbiome in health and disease.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Bacteria
/ Bacteria, Anaerobic - growth & development
/ Bacteria, Anaerobic - metabolism
/ Bacteroides thetaiotaomicron - metabolism
/ Coculture Techniques - methods
/ Genes
/ Humans
/ Hypoxia
/ Intestinal Mucosa - cytology
/ Intestinal Mucosa - metabolism
/ Intestinal Mucosa - microbiology
/ Medicine and Health Sciences
/ Microbiota (Symbiotic organisms)
/ Oxygen
