Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Single intestinal stem cells build crypt-villus structures in vitro without a cellular niche

ABSTRACT: The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. We have recently demonstrated the presence of approximately six cycling Lgr5+ stem cells at the bottoms of small intestinal crypts1. We have now established long-term culture conditions under which single crypts undergo multiple crypt fission events, whilst simultanously generating villus-like epithelial domains in which all differentiated cell types are present. Single sorted Lgr5+ stem cells can also initiate these crypt-villus organoids. Tracing experiments indicate that the Lgr5+ stem cell hierarchy is maintained in organoids. We conclude that intestinal crypt-villus units are self-organizing structures, which can be built from a single stem cell in the absence of a non-epithelial cellular niche. Keywords: expression profiling Freshly isolated small intestinal crypts from two mice were divided into two parts. RNA was directly isolated from one part (RNeasy Mini Kit, Qiagen), the other part was cultured for one week according to the conditions described in the associated paper, followed by RNA isolation. We prepared labeled cRNA following the manufacturer’s instruction (Agilent Technologies). Differentially labelled cRNA from small intestinal crypts and organoids were hybridised separately for the two mice on a 4X44k Agilent Whole Mouse Genome dual colour Microarrays (G4122F) in two dye swap experiments, resulting in four individual arrays.

ORGANISM(S): Mus musculus

SUBMITTER: Daniel Stange

PROVIDER: E-GEOD-14594 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.

Sato Toshiro T Vries Robert G RG Snippert Hugo J HJ van de Wetering Marc M Barker Nick N Stange Daniel E DE van Es Johan H JH Abo Arie A Kujala Pekka P Peters Peter J PJ Clevers Hans H

Nature 20090329 7244

The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. We have recently demonstrated the presence of about six cycling Lgr5(+) stem cells at the bottoms of small-intestinal crypts. Here we describe the establishment of long-term culture conditions under which single crypts undergo multiple crypt fission events, while simultanously generating villus-like epithelial domains in which all differentiated cell types are present. Single sorted Lgr5(+) stem cells can also i ...[more]

PMID: 19329995

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