Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche

Eugenio Sangiorgi, A Ootani, X Li, Qt Ho, H Ueno, S Toda, H Sugihara, K Fujimoto, Il Weissman, Mr Capecchi, Cj Kuo

Risultato della ricerca: Contributo in rivistaArticolo in rivistapeer review

503 Citazioni (Scopus)

Abstract

The in vitro analysis of intestinal epithelium has been hampered by a lack of suitable culture systems. Here we describe robust long-term methodology for small and large intestinal culture, incorporating an air-liquid interface and underlying stromal elements. These cultures showed prolonged intestinal epithelial expansion as sphere-like organoids with proliferation and multilineage differentiation. The Wnt growth factor family positively regulates proliferation of the intestinal epithelium in vivo. Accordingly, culture growth was inhibited by the Wnt antagonist Dickkopf-1 (Dkk1) and markedly stimulated by a fusion protein between the Wnt agonist R-spondin-1 and immunoglobulin Fc (RSpo1-Fc). Furthermore, treatment with the gamma-secretase inhibitor dibenzazepine and neurogenin-3 overexpression induced goblet cell and enteroendocrine cell differentiation, respectively, consistent with endogenous Notch signaling and lineage plasticity. Epithelial cells derived from both leucine-rich repeat-containing G protein-coupled receptor-5-positive (Lgr5(+)) and B lymphoma moloney murine leukemia virus insertion region homolog-1-positive (Bmi1(+)) lineages, representing putative intestinal stem cell (ISC) populations, were present in vitro and were expanded by treatment with RSpo1-Fc; this increased number of Lgr5(+) cells upon RSpo1-Fc treatment was subsequently confirmed in vivo. Our results indicate successful long-term intestinal culture within a microenvironment accurately recapitulating the Wnt- and Notch-dependent ISC niche.
Lingua originaleEnglish
pagine (da-a)701-706
Numero di pagine6
RivistaNature Medicine
Volume15
DOI
Stato di pubblicazionePubblicato - 2009

Keywords

  • Intestines
  • Nerve Tissue Proteins
  • Receptors, Notch
  • Signal Transduction
  • Stem Cell Niche
  • Thrombospondins
  • Tissue Culture Techniques
  • Wnt Proteins

Fingerprint

Entra nei temi di ricerca di 'Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche'. Insieme formano una fingerprint unica.

Cita questo