A Co-culture System for Intestinal Stem Cells (2009)

Undergraduate: Lieselotte Kreuk

Faculty Advisor: Susan Henning
Department: Biology

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Every 3-4 days our entire intestinal lining needs to be renewed because of the damage induced by luminal contents. This extremely high rate of epithelial turnover is achieved as a result of intestinal stem cells that reside in the intestinal crypts. In 2005, due to a breakthrough in the laboratory of my mentor Dr. Susan Henning, it was possible to isolate a viable fraction enriched for putative intestinal stem cells (PISC) and begin to understand their biology and develop their therapeutic potential. After being removed from their niche, I found that the PISC had limited viability in vitro, and they certainly did not proliferate or differentiate— two fundamental characteristics of stem cells. The long-term goal of my work is to recreate the intestinal niche by co-culturing the PISC with a potential critical element and observe proliferation. It is well known that a major cell type in the ISC niche is the pericryptal myofibroblast, which lies immediately subjacent to the overlying epithelial cells and is capable of secreting a wide range of critical factors. I co-cultured different characterized fibroblast lines with intestinal epithelial cells (IEC) in hopes of providing the important epithelial-mesenchymal interaction, enabling the specification of ISC during development. Preliminary studies included finding cell tracking methods that accurately quantify IEC viability and proliferation and the effects of different media conditions (i.e serum type and contentration).