Primary intestinal epithelial organoid culture

Tomohiro Mizutani, Hans Clevers

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

23 Citations (Scopus)

Abstract

The intestinal epithelium is known as one of the most regenerative tissues in our body. The lining of the intestine is composed of a single layer of epithelial cells generated by rapidly renewing stem cells residing at the crypt bottoms, resulting in a flow of cells to the villus tips. The stereotypical crypt–villus architecture makes the intestine an ideal model for stem cell research. Based on recent advances in research of stem cell niche signals in vivo, we have established an intestinal epithelial stem cell culture method. Under this culture condition, single Lgr5+ intestinal stem cells (ISCs) or isolated whole crypts efficiently expand into three-dimensional spherical structures recapitulating the intestinal crypt–villus organization. These organoids can be passaged weekly and maintained for years in culture. Moreover, they can be cryopreserved. As intestinal organoids recapitulate many aspects of the epithelial biology and are amenable to most, if not all, current experimental manipulations, they are widely used to study stem cell biology, cell fate determination, gene function, and disease mechanism.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages185-200
Number of pages16
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2171
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Colon
  • Crypt isolation
  • Intestinal epithelial stem cells
  • Lgr5
  • Mouse
  • Organoid culture
  • Small intestine

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