Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0

Gene expression signature of fast and slow cycling intestinal crypt base columnar cell populations


ABSTRACT: The identification of Lgr5 as an intestinal stem cell marker has made it possible to isolate and study primary stem cells from small intestine. Using the cell cycle specific expression og the mKi67 gene, we generated a novel Ki67-RFP knock-in allele which identifies dividing cells. Using Lgr5-GFP;Ki67-RFP mice, we isolated CBCs with distinct Wnt signaling levels and cell cycle features, and analyzed their global gene expression pattern using microarrays. We concluded that the cycling Lgr5hi stem cells exit the cell cycle in transition into the secretory lineage. Lgr5med Ki67low intermediate precursors reside in the zone of differentiation, resemble quiescent stem cells and generate the Dll1+ secretory precursors and the label retaining cells. Our findings support the cycling stem cell hypothesis and highlight the heterogeneity of early progenitors during lineage commitment. We used cell fractions of intestines from Lgr5-EGFP-ires-CreERT2 mice, expressing GFP under the control of the Lgr5 promoter, and Ki67-TagRFP mice where the RFP is fused to the C-terminus of the endogenous Ki67 gene. RNA was isolated from several FACS sorted cell populations of combinations expressing different levels of GFP and RFP: GFP high RFP high (Lgr5hi Ki67hi), GFP high RFP low (Lgr5hi Ki67low), GFP medium RFP high (Lgr5med Ki67high) and GFP medium RFP low (Lgr5med Ki67low). Purified RNA was processed, hybridized, and scanned according to the manufacturerM-bM-^@M-^Ys protocol and were hybridized on Affymetrix Mouse Gene ST 1.1 arrays).

ORGANISM(S): Mus musculus

SUBMITTER: Rogier Versteeg 

PROVIDER: E-GEOD-52813 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

altmetric image

Publications

Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele.

Basak Onur O   van de Born Maaike M   Korving Jeroen J   Beumer Joep J   van der Elst Stefan S   van Es Johan H JH   Clevers Hans H  

The EMBO journal 20140804 18


Cycling Lgr5+ stem cells fuel the rapid turnover of the adult intestinal epithelium. The existence of quiescent Lgr5+ cells has been reported, while an alternative quiescent stem cell population is believed to reside at crypt position +4. Here, we generated a novel Ki67RFP knock-in allele that identifies dividing cells. Using Lgr5-GFP;Ki67RFP mice, we isolated crypt stem and progenitor cells with distinct Wnt signaling levels and cell cycle features and generated their molecular signature using  ...[more]

Similar Datasets

2014-09-08 | GSE52813 | GEO
2012-08-15 | E-GEOD-36497 | biostudies-arrayexpress
2012-08-12 | E-GEOD-23672 | biostudies-arrayexpress
2010-11-29 | E-GEOD-25109 | biostudies-arrayexpress
2012-08-12 | E-GEOD-33948 | biostudies-arrayexpress
2013-04-25 | E-GEOD-46303 | biostudies-arrayexpress
2018-09-10 | E-MTAB-6891 | biostudies-arrayexpress
2012-09-27 | E-GEOD-39915 | biostudies-arrayexpress
2009-03-25 | E-GEOD-14201 | biostudies-arrayexpress
2013-08-01 | E-GEOD-49461 | biostudies-arrayexpress