PMID- 19676123 OWN - NLM STAT- MEDLINE DCOM- 20100217 LR - 20220409 IS - 1549-4918 (Electronic) IS - 1066-5099 (Print) IS - 1066-5099 (Linking) VI - 27 IP - 10 DP - 2009 Oct TI - Doublecortin and CaM kinase-like-1 and leucine-rich-repeat-containing G-protein-coupled receptor mark quiescent and cycling intestinal stem cells, respectively. PG - 2571-9 LID - 10.1002/stem.193 [doi] AB - It is thought that small intestinal epithelia (IE) undergo continuous self-renewal primarily due to their population of undifferentiated stem cells. These stem cells give rise to transit amplifying (daughter/progenitor) cells, which can differentiate into all mature cell types required for normal gut function. Identification of stem cells in IE is paramount to fully understanding this renewal process. One major obstacle in gastrointestinal stem cell biology has been the lack of definitive markers that identify small intestinal stem cells (ISCs). Here we demonstrate that the novel putative ISC marker doublecortin and CaM kinase-like-1 (DCAMKL-1) is predominantly expressed in quiescent cells in the lower two-thirds of intestinal crypt epithelium and in occasional crypt-based columnar cells (CBCs). In contrast, the novel putative stem cell marker leucine-rich-repeat-containing G-protein-coupled receptor (LGR5) is observed in rapidly cycling CBCs and in occasional crypt epithelial cells. Furthermore, functionally quiescent DCAMKL-1+ crypt epithelial cells retain bromo-deoxyuridine in a modified label retention assay. Moreover, we demonstrate that DCAMKL-1 is a cell surface expressing protein; DCAMKL-1+ cells, isolated from the adult mouse small intestine by fluorescence activated cell sorting, self-renew and ultimately form spheroids in suspension culture. These spheroids formed glandular epithelial structures in the flanks of athymic nude mice, which expressed multiple markers of gut epithelial lineage. Thus, DCAMKL-1 is a marker of quiescent ISCs and can be distinguished from the cycling stem/progenitors (LGR5+). Moreover, DCAMKL-1 can be used to isolate normal small intestinal stem cells and represents a novel research tool for regenerative medicine and cancer therapy. FAU - May, Randal AU - May R AD - Department of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA. FAU - Sureban, Sripathi M AU - Sureban SM FAU - Hoang, Nguyet AU - Hoang N FAU - Riehl, Terrence E AU - Riehl TE FAU - Lightfoot, Stan A AU - Lightfoot SA FAU - Ramanujam, Rama AU - Ramanujam R FAU - Wyche, James H AU - Wyche JH FAU - Anant, Shrikant AU - Anant S FAU - Houchen, Courtney W AU - Houchen CW LA - eng GR - K08 DK002822-05/DK/NIDDK NIH HHS/United States GR - DK-065887/DK/NIDDK NIH HHS/United States GR - R01 DK062265/DK/NIDDK NIH HHS/United States GR - R01 CA109269-09/CA/NCI NIH HHS/United States GR - R03 DK065887-04/DK/NIDDK NIH HHS/United States GR - K08 DK002822-02/DK/NIDDK NIH HHS/United States GR - R03 DK065887/DK/NIDDK NIH HHS/United States GR - R03 DK065887-03/DK/NIDDK NIH HHS/United States GR - K08 DK002822-01A1/DK/NIDDK NIH HHS/United States GR - R03 DK065887-02/DK/NIDDK NIH HHS/United States GR - K08 DK002822-04/DK/NIDDK NIH HHS/United States GR - DK-002822/DK/NIDDK NIH HHS/United States GR - R03 DK065887-01/DK/NIDDK NIH HHS/United States GR - K08 DK002822/DK/NIDDK NIH HHS/United States GR - R01 CA135559-03/CA/NCI NIH HHS/United States GR - R01 CA109269/CA/NCI NIH HHS/United States GR - R01 CA135559/CA/NCI NIH HHS/United States GR - R01 DK062265-08/DK/NIDDK NIH HHS/United States GR - K08 DK002822-03/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. PL - England TA - Stem Cells JT - Stem cells (Dayton, Ohio) JID - 9304532 RN - 0 (Antigens, Surface) RN - 0 (Biomarkers) RN - 0 (Lgr5 protein, mouse) RN - 0 (Receptors, G-Protein-Coupled) RN - EC 2.7.1.11 (Doublecortin-Like Kinases) RN - EC 2.7.11.1 (Dclk1 protein, mouse) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - G34N38R2N1 (Bromodeoxyuridine) SB - IM MH - Animals MH - Antigens, Surface/analysis/metabolism MH - Biomarkers/analysis/metabolism MH - Bromodeoxyuridine MH - Cell Cycle/physiology MH - Cell Division/physiology MH - Cell Line, Tumor MH - Cell Lineage/physiology MH - Cell Proliferation MH - Doublecortin-Like Kinases MH - Flow Cytometry MH - Humans MH - Intestinal Mucosa/cytology/*metabolism MH - Mice MH - Mice, Inbred C57BL MH - Mice, Nude MH - Protein Serine-Threonine Kinases/analysis/*metabolism MH - Receptors, G-Protein-Coupled/analysis/*metabolism MH - Spheroids, Cellular MH - Stem Cells/cytology/*metabolism PMC - PMC3049723 MID - NIHMS275972 EDAT- 2009/08/14 09:00 MHDA- 2010/02/18 06:00 PMCR- 2011/03/07 CRDT- 2009/08/14 09:00 PHST- 2009/08/14 09:00 [entrez] PHST- 2009/08/14 09:00 [pubmed] PHST- 2010/02/18 06:00 [medline] PHST- 2011/03/07 00:00 [pmc-release] AID - 10.1002/stem.193 [doi] PST - ppublish SO - Stem Cells. 2009 Oct;27(10):2571-9. doi: 10.1002/stem.193.