Primary mouse small intestinal epithelial cell cultures

Toshiro Sato, Hans Clevers

Research output: Chapter in Book/Report/Conference proceedingChapter

111 Citations (Scopus)

Abstract

The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. We have recently shown that Lgr5 (Leucine-rich repeat-containing G protein-coupled receptor) is expressed in intestinal stem cells by an in vivo genetic lineage tracing strategy. In the past, extensive efforts have been made to establish primary small intestinal culture systems. However, no defined, reproducible and robust culture system had been developed. To establish such a system, we screened for optimal growth factor combinations based on genetic evidence of self-renewal regulation, differentiation, and carcinogenesis of intestinal stem cells. Here, we describe methods that we have established for the isolation and culture of primary small intestinal epithelial stem cells. In this culture system, isolated crypts form organoid structures with a histological hierarchy recapitulating in vivo small intestinal epithelium. Single isolated Lgr5+ intestinal stem cells also form these organoid structures, in which stem cells are maintained by self-renewal and give rise to all lineages of the intestinal epithelium. This culture system is particularly useful for studying the regulation of intestinal stem cell self-renewal and differentiation.

Original languageEnglish
Title of host publicationEpithelial Cell Culture Protocols: Second Edition
Pages319-328
Number of pages10
Volume945
DOIs
Publication statusPublished - 2013

Publication series

NameMethods in Molecular Biology
Volume945
ISSN (Print)10643745

Fingerprint

Stem Cells
Intestinal Mucosa
Cell Culture Techniques
Epithelial Cells
Organoids
G-Protein-Coupled Receptors
Leucine
Mammals
Intercellular Signaling Peptides and Proteins
Carcinogenesis
Cell Self Renewal

Keywords

  • Confocal immunofluorescence
  • Flow cytometry
  • Intestinal stem cells
  • Lgr5
  • Small intestinal crypts
  • Wnt signal

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Sato, T., & Clevers, H. (2013). Primary mouse small intestinal epithelial cell cultures. In Epithelial Cell Culture Protocols: Second Edition (Vol. 945, pp. 319-328). (Methods in Molecular Biology; Vol. 945). https://doi.org/10.1007/978-1-62703-125-7_19

Primary mouse small intestinal epithelial cell cultures. / Sato, Toshiro; Clevers, Hans.

Epithelial Cell Culture Protocols: Second Edition. Vol. 945 2013. p. 319-328 (Methods in Molecular Biology; Vol. 945).

Research output: Chapter in Book/Report/Conference proceedingChapter

Sato, T & Clevers, H 2013, Primary mouse small intestinal epithelial cell cultures. in Epithelial Cell Culture Protocols: Second Edition. vol. 945, Methods in Molecular Biology, vol. 945, pp. 319-328. https://doi.org/10.1007/978-1-62703-125-7_19
Sato T, Clevers H. Primary mouse small intestinal epithelial cell cultures. In Epithelial Cell Culture Protocols: Second Edition. Vol. 945. 2013. p. 319-328. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-125-7_19
Sato, Toshiro ; Clevers, Hans. / Primary mouse small intestinal epithelial cell cultures. Epithelial Cell Culture Protocols: Second Edition. Vol. 945 2013. pp. 319-328 (Methods in Molecular Biology).
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