A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction.
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...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2024-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0300666 |
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| author | Tatiana Y Fofanova Umesh C Karandikar Jennifer M Auchtung Reid L Wilson Antonio J Valentin Robert A Britton K Jane Grande-Allen Mary K Estes Kristi Hoffman Sashirekha Ramani Christopher J Stewart Joseph F Petrosino |
| author_facet | Tatiana Y Fofanova Umesh C Karandikar Jennifer M Auchtung Reid L Wilson Antonio J Valentin Robert A Britton K Jane Grande-Allen Mary K Estes Kristi Hoffman Sashirekha Ramani Christopher J Stewart Joseph F Petrosino |
| author_sort | Tatiana Y Fofanova |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-390d9f33524946f68a4b66b3442bcb4d |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-390d9f33524946f68a4b66b3442bcb4d2025-08-20T03:51:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-01197e030066610.1371/journal.pone.0300666A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction.Tatiana Y FofanovaUmesh C KarandikarJennifer M AuchtungReid L WilsonAntonio J ValentinRobert A BrittonK Jane Grande-AllenMary K EstesKristi HoffmanSashirekha RamaniChristopher J StewartJoseph F PetrosinoMechanistic 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.https://doi.org/10.1371/journal.pone.0300666 |
| spellingShingle | Tatiana Y Fofanova Umesh C Karandikar Jennifer M Auchtung Reid L Wilson Antonio J Valentin Robert A Britton K Jane Grande-Allen Mary K Estes Kristi Hoffman Sashirekha Ramani Christopher J Stewart Joseph F Petrosino A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. PLoS ONE |
| title | A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. |
| title_full | A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. |
| title_fullStr | A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. |
| title_full_unstemmed | A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. |
| title_short | A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction. |
| title_sort | novel system to culture human intestinal organoids under physiological oxygen content to study microbial host interaction |
| url | https://doi.org/10.1371/journal.pone.0300666 |
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