Primary mouse small intestinal epithelial cell cultures

Toshiro Sato, Hans Clevers

研究成果: Chapter

170 被引用数 (Scopus)

抄録

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.

本文言語English
ホスト出版物のタイトルEpithelial Cell Culture Protocols
ホスト出版物のサブタイトルSecond Edition
編集者Scott Randell, Leslie Fulcher
ページ319-328
ページ数10
DOI
出版ステータスPublished - 2013

出版物シリーズ

名前Methods in Molecular Biology
945
ISSN(印刷版)1064-3745

ASJC Scopus subject areas

  • 分子生物学
  • 遺伝学

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