Advertisement

Organoids pp 183-194 | Cite as

Intestinal Organoids as a Novel Tool to Study Microbes–Epithelium Interactions

  • Giulia NigroEmail author
  • Melissa Hanson
  • Cindy Fevre
  • Marc Lecuit
  • Philippe J. Sansonetti
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1576)

Abstract

The gut, particularly the colon, is the host of approximately 1000 bacterial species, the so-called gut microbiota. The relationship between the gut microbiota and the host is symbiotic and mutualistic, influencing many aspects of the biology of the host. This homeostatic balance can be disrupted by enteric pathogens, such as Shigella flexneri or Listeria monocytogenes, which are able to invade the epithelial layer and consequently subvert physiological functions. To study the host–microbe interactions in vitro, the crypt culture model, known as intestinal organoids, is a powerful tool. Intestinal organoids provide a model in which to examine the response of the epithelium, particularly the response of intestinal stem cells, to the presence of bacteria. Furthermore, the organoid model enables the study of pathogens during the early steps of enteric pathogen invasion.

Here, we describe methods that we have established to study the cellular microbiology of symbiosis between the gut microbiota and host intestinal surface and secondly the disruption of host homeostasis due to an enteric pathogen.

Keywords:

Intestinal organoids Host–microbe interactions Listeria monocytogenes Bacterial products Flow cytometry Microinjection 

References

  1. 1.
    Sansonetti PJ (2004) War and peace at mucosal surfaces. Nat Rev Immunol 4:953–964CrossRefGoogle Scholar
  2. 2.
    Sato T, Vries RG, Snippert HJ et al (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459:262–265CrossRefGoogle Scholar
  3. 3.
    Nigro G, Rossi R, Commere P-H et al (2014) The cytosolic bacterial peptidoglycan sensor Nod2 affords stem cell protection and links microbes to gut epithelial regeneration. Cell Host Microbe 15:792–798CrossRefGoogle Scholar
  4. 4.
    Bonazzi M, Lecuit M, Cossart P (2009) Listeria monocytogenes internalin and E-cadherin: from structure to pathogenesis. Cell Microbiol 11:693–702CrossRefGoogle Scholar
  5. 5.
    Nikitas G, Deschamps C, Disson O et al (2011) Transcytosis of Listeria monocytogenes across the intestinal barrier upon specific targeting of goblet cell accessible E-cadherin. J Exp Med 208:2263–2277CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 2.5 International License (http://creativecommons.org/licenses/by-nc/2.5/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

The images or other third party material in this chapter are included in the chapter's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the chapter's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

Authors and Affiliations

  • Giulia Nigro
    • 1
    Email author
  • Melissa Hanson
    • 2
  • Cindy Fevre
    • 2
  • Marc Lecuit
    • 2
  • Philippe J. Sansonetti
    • 1
  1. 1.Molecular Microbial Pathogenesis Unit, Inserm U1202Institut PasteurParisFrance
  2. 2.Biology of Infection Unit, Inserm U1117Institut PasteurParisFrance

Personalised recommendations