A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction

Authors

Tatiana Y. Fofanova, Baylor College of Medicine
Umesh C. Karandikar, Baylor College of Medicine
Jennifer M. Auchtung, Baylor College of Medicine, University of Nebraska-LincolnFollow
Reid L. Wilson, Rice University, Baylor College of Medicine
Antonio J. Valentin, Baylor College of Medicine
Robert A. Britton, Baylor College of Medicine
K. Jane Grande-Allen, Rice University
Mary K. Estes, Baylor College of Medicine
Kristi Hoffman, Baylor College of Medicine
Sashirekha Ramani, Baylor College of Medicine
Christopher J. Stewart, Baylor College of Medicine, Newcastle University
Joseph F. Petrosino, Baylor College of Medicine

Date of this Version

3-1-2024

Document Type

Article

Citation

Fofanova TY, Karandikar UC, Auchtung JM, Wilson RL, Valentin AJ, Britton RA, et al. (2024) A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. PLoS ONE 19(7): e0300666. https://doi.org/10.1371/journal. pone.0300666

Comments

Open access.

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.