Establishment and Culture of Human Intestinal Organoids Derived from Adult Stem Cells

Cayetano Pleguezuelos-Manzano, Jens Puschhof, Stieneke van den Brink, Veerle Geurts, Joep Beumer, Hans Clevers

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86 Citations (Scopus)


Human intestinal organoids derived from adult stem cells are miniature ex vivo versions of the human intestinal epithelium. Intestinal organoids are useful tools for the study of intestinal physiology as well as many disease conditions. These organoids present numerous advantages compared to immortalized cell lines, but working with them requires dedicated techniques. The protocols described in this article provide a basic guide to establishment and maintenance of human intestinal organoids derived from small intestine and colon biopsies. Additionally, this article provides an overview of several downstream applications of human intestinal organoids.

Original languageEnglish
Article numbere106
JournalCurrent Protocols in Immunology
Issue number1
Publication statusPublished - 1 Sept 2020


  • adult stem cells
  • human intestinal organoids
  • organoid cryopreservation
  • organoid culture establishment
  • organoid differentiation
  • organoid immunofluorescence
  • organoid passage
  • single-cell clonal organoid culture
  • specialized organoid reagents


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