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    Isolation and Characterization of Quiescent Intestinal Stem Cells

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    Scoville_ku_0099D_11023_DATA_1.pdf (193.9Mb)
    Issue Date
    2010-06-10
    Author
    Scoville, David
    Publisher
    University of Kansas
    Format
    167 pages
    Type
    Dissertation
    Degree Level
    Ph.D.
    Discipline
    Pathology & Laboratory Medicine
    Rights
    This item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author.
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    Abstract
    Critical to the identification of putative stem cell populations is the ability to evalu-ate their functional potential. Within this work I describe the development of a robust culture system which enables regeneration of the intestinal epithelium from isolated crypts and single intestinal stem cells. To discriminate putative quiescent intestinal stem cells (ISCs) from rapidly cycling Lgr5+ ISCs a tetracycline inducible H2BGFP labeling strategy was employed. Distinct populations of label and non-label retaining cells were separated via fluorescent activated cell sorting (FACS). Analysis of stem cell potential, using the intesti-nal in vitro culture system developed herein, revealed that CD166high label retaining cells (LRCs) and non-LRCs contain functional intestinal stem cells (ISCs). Gene expression analysis of these populations confirmed the expression of stem cell genes including Lgr5. Coupled with the identification of long term BrdU retaining Lgr5+ cells, this work demon-strates the existence of both active and quiescent Lgr5+ ISCs.
    URI
    http://hdl.handle.net/1808/7385
    Collections
    • Dissertations [4474]
    • KU Med Center Dissertations and Theses [464]

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    785-864-8983
    KU Libraries
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    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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