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Versatile human in vitro triple coculture model coincubated with adhered gut microbes reproducibly mimics pro-inflammatory host-microbe interactions in the colon

(2021) FASEB JOURNAL. 35(12).
Author
Organization
Abstract
The colonic epithelial barrier is vital to preserve gut and host health by maintaining the immune homeostasis between host and microbes. The mechanisms underlying beneficial or harmful host-microbe interactions are poorly understood and impossible to study in vivo given the limited accessibility and ethical constraints. Moreover, existing in vitro models lack the required cellular complexity for the routine, yet profound, analysis of the intricate interplay between different types of host and microbial cells. We developed and characterized a broadly applicable, easy-to-handle in vitro triple coculture model that combines chemically-induced macrophage-like, goblet and epithelial cells covered by a mucus layer, which can be coincubated with complex human-derived gut microbiota samples for 16 h. Comparison with a standard epithelial monolayer model revealed that triple cocultures produce thicker mucus layers, morphologically organize in a network and upon exposure to human-derived gut microbiota samples, respond via pro-inflammatory cytokine production. Both model systems, however, were not suffering from cytotoxic stress or different microbial loads, indicating that the obtained endpoints were caused by the imposed conditions. Addition of the probiotic Lactobacillus rhamnosus GG to assess its immunomodulating capacity in the triple coculture slightly suppressed pro-inflammatory cytokine responses, based on transcriptomic microarray analyses. TNF conditioning of the models prior to microbial exposure did not cause shifts in cytokines, suggesting a strong epithelial barrier in which TNF did not reach the basolateral side. To conclude, the triple coculture model is tolerable towards manipulations and allows to address mechanistic host-microbe research questions in a stable in vitro environment.
Keywords
Genetics, Molecular Biology, Biochemistry, Biotechnology, coculture techniques, epithelial cells, gastrointestinal microbiome, gene expression profiling, host microbial interactions, LACTOBACILLUS-RHAMNOSUS GG, EPITHELIAL-CELL LINES, NECROSIS-FACTOR-ALPHA, GENE-EXPRESSION, HUMAN INTESTINE, CACO-2, INTERLEUKIN-8, BARRIER, CACO-2/HT29-MTX, ORGANOIDS

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Citation

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MLA
Beterams, Annelore, et al. “Versatile Human in Vitro Triple Coculture Model Coincubated with Adhered Gut Microbes Reproducibly Mimics Pro-Inflammatory Host-Microbe Interactions in the Colon.” FASEB JOURNAL, vol. 35, no. 12, 2021, doi:10.1096/fj.202101135r.
APA
Beterams, A., De Paepe, K., Maes, L., Wise, I. J., De Keersmaecker, H., Rajkovic, A., … Calatayud Arroyo, M. (2021). Versatile human in vitro triple coculture model coincubated with adhered gut microbes reproducibly mimics pro-inflammatory host-microbe interactions in the colon. FASEB JOURNAL, 35(12). https://doi.org/10.1096/fj.202101135r
Chicago author-date
Beterams, Annelore, Kim De Paepe, Laure Maes, India Jane Wise, Herlinde De Keersmaecker, Andreja Rajkovic, Debby Laukens, Tom Van de Wiele, and Marta Calatayud Arroyo. 2021. “Versatile Human in Vitro Triple Coculture Model Coincubated with Adhered Gut Microbes Reproducibly Mimics Pro-Inflammatory Host-Microbe Interactions in the Colon.” FASEB JOURNAL 35 (12). https://doi.org/10.1096/fj.202101135r.
Chicago author-date (all authors)
Beterams, Annelore, Kim De Paepe, Laure Maes, India Jane Wise, Herlinde De Keersmaecker, Andreja Rajkovic, Debby Laukens, Tom Van de Wiele, and Marta Calatayud Arroyo. 2021. “Versatile Human in Vitro Triple Coculture Model Coincubated with Adhered Gut Microbes Reproducibly Mimics Pro-Inflammatory Host-Microbe Interactions in the Colon.” FASEB JOURNAL 35 (12). doi:10.1096/fj.202101135r.
Vancouver
1.
Beterams A, De Paepe K, Maes L, Wise IJ, De Keersmaecker H, Rajkovic A, et al. Versatile human in vitro triple coculture model coincubated with adhered gut microbes reproducibly mimics pro-inflammatory host-microbe interactions in the colon. FASEB JOURNAL. 2021;35(12).
IEEE
[1]
A. Beterams et al., “Versatile human in vitro triple coculture model coincubated with adhered gut microbes reproducibly mimics pro-inflammatory host-microbe interactions in the colon,” FASEB JOURNAL, vol. 35, no. 12, 2021.
@article{8725830,
  abstract     = {{The colonic epithelial barrier is vital to preserve gut and host health by maintaining the immune homeostasis between host and microbes. The mechanisms underlying beneficial or harmful host-microbe interactions are poorly understood and impossible to study in vivo given the limited accessibility and ethical constraints. Moreover, existing in vitro models lack the required cellular complexity for the routine, yet profound, analysis of the intricate interplay between different types of host and microbial cells. We developed and characterized a broadly applicable, easy-to-handle in vitro triple coculture model that combines chemically-induced macrophage-like, goblet and epithelial cells covered by a mucus layer, which can be coincubated with complex human-derived gut microbiota samples for 16 h. Comparison with a standard epithelial monolayer model revealed that triple cocultures produce thicker mucus layers, morphologically organize in a network and upon exposure to human-derived gut microbiota samples, respond via pro-inflammatory cytokine production. Both model systems, however, were not suffering from cytotoxic stress or different microbial loads, indicating that the obtained endpoints were caused by the imposed conditions. Addition of the probiotic Lactobacillus rhamnosus GG to assess its immunomodulating capacity in the triple coculture slightly suppressed pro-inflammatory cytokine responses, based on transcriptomic microarray analyses. TNF conditioning of the models prior to microbial exposure did not cause shifts in cytokines, suggesting a strong epithelial barrier in which TNF did not reach the basolateral side. To conclude, the triple coculture model is tolerable towards manipulations and allows to address mechanistic host-microbe research questions in a stable in vitro environment.}},
  articleno    = {{e21992}},
  author       = {{Beterams, Annelore and De Paepe, Kim and Maes, Laure and Wise, India Jane and De Keersmaecker, Herlinde and Rajkovic, Andreja and Laukens, Debby and Van de Wiele, Tom and Calatayud Arroyo, Marta}},
  issn         = {{0892-6638}},
  journal      = {{FASEB JOURNAL}},
  keywords     = {{Genetics,Molecular Biology,Biochemistry,Biotechnology,coculture techniques,epithelial cells,gastrointestinal microbiome,gene expression profiling,host microbial interactions,LACTOBACILLUS-RHAMNOSUS GG,EPITHELIAL-CELL LINES,NECROSIS-FACTOR-ALPHA,GENE-EXPRESSION,HUMAN INTESTINE,CACO-2,INTERLEUKIN-8,BARRIER,CACO-2/HT29-MTX,ORGANOIDS}},
  language     = {{eng}},
  number       = {{12}},
  pages        = {{19}},
  title        = {{Versatile human in vitro triple coculture model coincubated with adhered gut microbes reproducibly mimics pro-inflammatory host-microbe interactions in the colon}},
  url          = {{http://dx.doi.org/10.1096/fj.202101135r}},
  volume       = {{35}},
  year         = {{2021}},
}

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