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Methods Mol Biol. 2016 Aug 31. [Epub ahead of print]

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

Author information

1
Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK. jf8@sanger.ac.uk.
2
University of Nottingham, Nottingham, CB10 1SA, UK.
3
Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
4
Wellcome Trust-Medical Research Council Stem Cell Institute, Anne McLaren Laboratory, Department of Surgery, University of Cambridge, Cambridge, UK.

Abstract

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).

KEYWORDS:

Differentiation; Host–pathogen interactions; Intestinal epithelium; Microinjection; Organoids; hIPSCs

PMID:
27576565
DOI:
10.1007/7651_2016_7

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