1. A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction.
- Author
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Fofanova, Tatiana Y., Karandikar, Umesh C., Auchtung, Jennifer M., Wilson, Reid L., Valentin, Antonio J., Britton, Robert A., Grande-Allen, K. Jane, Estes, Mary K., Hoffman, Kristi, Ramani, Sashirekha, Stewart, Christopher J., and Petrosino, Joseph F.
- Subjects
INTESTINAL mucosa ,ANAEROBIC bacteria ,ORGANOIDS ,ANAEROBIC microorganisms ,OXYGEN ,EPITHELIAL cells ,BACTEROIDES fragilis ,INTESTINES ,SMALL intestine - Abstract
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 vivo. 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. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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