Document Detail


CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice.
MedLine Citation:
PMID:  22723265     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24(+) fraction that contained the majority of Lgr5-EGFP(+) putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24(+)EpCAM(+) fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase-like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies.
Authors:
Jeffrey B King; Richard J von Furstenberg; Brian J Smith; Kirk K McNaughton; Joseph A Galanko; Susan J Henning
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-06-21
Journal Detail:
Title:  American journal of physiology. Gastrointestinal and liver physiology     Volume:  303     ISSN:  1522-1547     ISO Abbreviation:  Am. J. Physiol. Gastrointest. Liver Physiol.     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-08-16     Completed Date:  2012-10-25     Revised Date:  2013-08-18    
Medline Journal Info:
Nlm Unique ID:  100901227     Medline TA:  Am J Physiol Gastrointest Liver Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  G443-52     Citation Subset:  IM    
Affiliation:
Department of Medicine, University of North Carolina at Chapel Hill, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, CD24 / metabolism*
Antigens, Neoplasm / metabolism
Biological Markers / metabolism
Cell Adhesion Molecules / metabolism
Cell Proliferation
Cell Separation / methods*
Colon / cytology,  immunology*
Epithelial Cells / immunology*
Flow Cytometry*
Gene Expression Regulation
Green Fluorescent Proteins / biosynthesis,  genetics
Immunohistochemistry
Intestinal Mucosa / cytology,  immunology*
Mice
Mice, Inbred C57BL
Mice, Transgenic
Microscopy, Confocal
RNA, Messenger / metabolism
Receptors, G-Protein-Coupled / genetics,  metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells / immunology*
Transcription, Genetic
Grant Support
ID/Acronym/Agency:
P30 CA-06086/CA/NCI NIH HHS; P30-DK-034987/DK/NIDDK NIH HHS; T32-DK-007737/DK/NIDDK NIH HHS; U01-DK-085547-03S1/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD24; 0/Antigens, Neoplasm; 0/Biological Markers; 0/Cd24a protein, mouse; 0/Cell Adhesion Molecules; 0/Lgr5 protein, mouse; 0/RNA, Messenger; 0/Receptors, G-Protein-Coupled; 0/enhanced green fluorescent protein; 0/tumor-associated antigen GA733; 147336-22-9/Green Fluorescent Proteins
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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