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Derivation of Intestinal Organoids from Human Induced Pluripotent Stem Cells for Use as an Infection System

  • Jessica L. ForbesterEmail author
  • Nicholas Hannan
  • Ludovic Vallier
  • Gordon Dougan
Part of the Methods in Molecular Biology book series


Intestinal human organoids (iHOs) provide an effective system for studying the intestinal epithelium and its interaction with various stimuli. By using combinations of different signaling factors, human induced pluripotent stem cells (hIPSCs) can be driven to differentiate down the intestinal lineage. Here, we describe the process for this differentiation, including the derivation of hindgut from hIPSCs, embedding hindgut into a pro-intestinal culture system and passaging the resulting iHOs. We then describe how to carry out microinjections to introduce bacteria to the apical side of the intestinal epithelial cells (IECs).


Organoids hIPSCs Intestinal epithelium Differentiation Microinjection Host–pathogen interactions 



This work was supported by the Medical Research Council and the Wellcome Trust. We thank David Goulding for providing TEM images.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jessica L. Forbester
    • 1
    Email author
  • Nicholas Hannan
    • 2
  • Ludovic Vallier
    • 1
    • 3
  • Gordon Dougan
    • 1
  1. 1.Wellcome Trust Sanger InstituteHinxton, CambridgeUK
  2. 2.University of NottinghamNottinghamUK
  3. 3.Wellcome Trust-Medical Research Council Stem Cell Institute, Anne McLaren Laboratory, Department of SurgeryUniversity of CambridgeCambridgeUK

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