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Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

DOI:
10.3791/59478-v

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10:59 min

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May 12, 2019

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, , , , ,
1Wellcome Trust Sanger Institute, 2Department of Medicine,University of Cambridge, 3University of Cardiff

Capitoli

  • 00:04Titolo
  • 00:38Differentiation from iPSCs to the Hindgut
  • 01:10Embedding of the Hindgut in Basement Membrane Matrix
  • 03:13Maintenance, Passage, and Pre-stimulation of iHOs with rhIL-22
  • 05:28Microinjection of iHOs and Intracellular Invasion Assays
  • 09:19Results: Studying Epithelial Cell Protection Against Pathogens Using hiPSC-derived iHOs
  • 10:05Conclusion

Summary

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Traduzione automatica

Human induced pluripotent stem cell (hiPSC)-derived intestinal organoids offer exciting opportunities to model enteric diseases in vitro. We demonstrate the differentiation of hiPSCs into intestinal organoids (iHOs), the stimulation of these iHOs with cytokines, and the microinjection of Salmonella Typhimurium into the iHO lumen, enabling the study of an epithelial invasion by this pathogen.

Tags

Human Induced Pluripotent Stem CellsIntestinal OrganoidsEpithelial Cell ProtectionSalmonellaPathogensMicro-injectionDifferentiationRPMI-B27 MediaCHIR99021Retinoic AcidBasement Membrane MatrixCollagenaseOrganoid Base Growth MediaCentrifugation24-well Plate

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