title: Derivation of Intestinal Organoids from Human Induced Pluripotent Stem Cells for Use as an Infection System.
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).
authors: Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK. jf8@sanger.ac.uk.
authors: Jessica L
authors: JL
authors: Forbester
authors: University of Nottingham, Nottingham, CB10 1SA, UK.
authors: Nicholas
authors: N
authors: Hannan
authors: Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.|Wellcome Trust-Medical Research Council Stem Cell Institute, Anne McLaren Laboratory, Department of Surgery, University of Cambridge, Cambridge, UK.
authors: Ludovic
authors: L
authors: Vallier
authors: Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
authors: Gordon
authors: G
authors: Dougan
mesh_terms: D001419:Bacteria / Q000254:growth & development*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D054884:Host-Pathogen Interactions; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000382:microbiology; D007422:Intestines / Q000166:cytology* / Q000382:microbiology; D009940:Organoids / Q000166:cytology* / Q000382:microbiology
keywords: Differentiation; Host–pathogen interactions; Intestinal epithelium; Microinjection; Organoids; hIPSCs