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Series GSE52402 Query DataSets for GSE52402
Status Public on Dec 20, 2013
Title Intestinal subepithelial myofibroblasts support the growth of intestinal epithelial stem cells
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Intestinal epithelial stem cells (ISCs) are the focus of recent intense study. Current in vitro models rely on supplementation with the Wnt agonist R-spondin1 to support robust growth, ISC self-renewal, and differentiation. Intestinal subepithelial myofibroblasts (ISEMFs) are important supportive cells within the ISC niche. We hypothesized that co-culture with ISEMF enhances the growth of ISCs in vitro and allows for their successful in vivo implantation and engraftment. ISC-containing small intestinal crypts, FACS-sorted single ISCs, and ISEMFs were procured from C57BL/6 mice. Crypts and single ISCs were grown in vitro into enteroids, in the presence or absence of ISEMFs. ISEMFs enhanced the growth of intestinal epithelium in vitro in a proximity-dependent fashion, with co-cultures giving rise to larger enteroids than monocultures. Co-culture of ISCs with supportive ISEMFs relinquished the requirement of exogenous R-spondin1 to sustain long-term growth and differentiation of ISCs. Mono- and co-cultures were implanted subcutaneously in syngeneic mice. Co-culture with ISEMFs proved necessary for successful in vivo engraftment and proliferation of enteroids; implants without ISEMFs did not survive. ISEMF whole transcriptome sequencing and qPCR demonstrated high expression of specific R-spondins, well-described Wnt agonists that supports ISC growth. Specific non-supportive ISEMF populations had reduced expression of R-spondins. The addition of ISEMFs in intestinal epithelial culture therefore recapitulates a critical element of the intestinal stem cell niche and allows for its experimental interrogation and biodesign-driven manipulation.
 
Overall design Two samples of intestinal subepithelial myofibroblasts were used in this study.
 
Contributor(s) Lei N, Jabaji Z, Wang J, Joshi VS, Brinkley GJ, Khalil H, Wang F, Jaroszewicz A, Pellegrini M, Li L, Lewis M, Stelzner M, Dunn JC, Martin MG
Citation(s) 24400106
Submission date Nov 15, 2013
Last update date May 15, 2019
Contact name Martin G Martin
E-mail(s) mmartin@mednet.ucla.edu
Organization name University of California Los Angeles
Department Pediatrics Gastroenterology
Street address BOX 951752, 12-383 MDCC
City Los Angeles
State/province CA
ZIP/Postal code 90095
Country USA
 
Platforms (1)
GPL13112 Illumina HiSeq 2000 (Mus musculus)
Samples (2)
GSM1264934 B19 - Supportive ISEMF
GSM1264935 B20 - Non-Supportive ISEMF
Relations
BioProject PRJNA227798
SRA SRP033020

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE52402_B19P16_B20P3_RPKM.txt.gz 2.5 Mb (ftp)(http) TXT
GSE52402_Wnt_pathway_RPKM.txt.gz 53.6 Kb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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