Abstract
Introduction: Intestinal stem cells can form spheroids when they are cultured within Matrigel supplemented with the Wnt agonist R-spondin that interacts with the Lgr5 receptor. A critical barrier to translate this methodology to cell-based therapy is the ability to generate a sheet of intestinal epithelial cells using well-defined extracellular matrix. We hypothesize that collagen gel will induce the formation of such epithelial cell sheets and eliminate the need for Matrigel. Methods: Intestinal crypts were isolated from transgenic GFP mice between 4-6 weeks of age using EDTA chelation.
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