Li, LinhengSoares, MichaelScoville, David2011-04-252011-04-252010-06-102010http://dissertations.umi.com/ku:11023https://hdl.handle.net/1808/7385Critical 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.167 pagesen-USThis item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author.Cell biologyIntestinal culture systemLabel retaining stem cellsQuiescent intestinal stem cellsDissertationopenAccess