Doublecortin and CaM Kinase-like-1 and Leucine-Rich-Repeat-Containing G-Protein-Coupled Receptor Mark Quiescent and Cycling Intestinal Stem Cells, Respectively§

Authors

  • Randal May,

    1. Departments of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    2. Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma 73104
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  • Sripathi M. Sureban,

    1. Departments of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    2. Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma 73104
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  • Nguyet Hoang,

    1. Departments of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
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  • Terrence E. Riehl,

    1. Department of Internal Medicine, Division of Gastroenterology, Washington University School of Medicine, St. Louis, Missouri 63110
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  • Stan A. Lightfoot,

    1. Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    2. Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma 73104
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  • Rama Ramanujam,

    1. PanCagen Inc., Oklahoma City, Oklahoma 73104
    2. ADNA Inc., Dublin, Ohio 43017
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  • James H. Wyche,

    1. PanCagen Inc., Oklahoma City, Oklahoma 73104
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  • Shrikant Anant,

    1. Departments of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    2. Cell Biology andThe University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    3. OU Cancer Institute, Oklahoma City, Oklahoma 73104
    4. PanCagen Inc., Oklahoma City, Oklahoma 73104
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  • Courtney W. Houchen

    Corresponding author
    1. Departments of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
    2. OU Cancer Institute, Oklahoma City, Oklahoma 73104
    3. Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma 73104
    4. PanCagen Inc., Oklahoma City, Oklahoma 73104
    • Department of Medicine, Digestive Diseases and Nutrition Section, University of Oklahoma Health Sciences Center, 920 Stanton L. Young Boulevard, WP 1360, Oklahoma City, OK 73104
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    • Telephone: (405) 271-2175; Fax: (405) 271-5450


  • Author contributions: R.M., S.M.S.: conception and design, collection and assembly of data, data analysis and integration, manuscript writing, and final approval of the manuscript; T.E.R.: manuscript writing and final approval of the manuscript; S.A.L., R.R., J.W.H.: data analysis and integration and final approval of the manuscript; S.A.: conception and design, data analysis, and integration and final approval of the manuscript; C.W.H.: conception and design, financial support, collection and assembly of data, data analysis and integration, manuscript writing, and final approval of the manuscript.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLS EXPRESS August 12, 2009.

Abstract

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. STEM CELLS 2009;27:2571–2579

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