Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+ stem cell

Building on their recent work establishing long-term cultures from Lgr5 + cells of the small intestine and stomach, Shiro Yui and colleagues describe a new colonic stem cell culture and expansion method generating organoids from single Lgr5 + stem cells. The study provides proof of principle that the cultured Lgr5 + cells can be used for stem cell therapy to repair superficially damaged epithelium in a dextran sulfate sodium (DSS)-induced acute colitis mouse model. Adult stem-cell therapy holds promise for the treatment of gastrointestinal diseases. Here we describe methods for long-term expansion of colonic stem cells positive for leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5 + cells) in culture. To test the transplantability of these cells, we reintroduced cultured GFP + colon organoids into superficially damaged mouse colon. The transplanted donor cells readily integrated into the mouse colon, covering the area that lacked epithelium as a result of the introduced damage in recipient mice. At 4 weeks after transplantation, the donor-derived cells constituted a single-layered epithelium, which formed self-renewing crypts that were functionally and histologically normal. Moreover, we observed long-term (>6 months) engraftment with transplantation of organoids derived from a single Lgr5 + colon stem cell after extensive in vitro expansion. These data show the feasibility of colon stem-cell therapy based on the in vitro expansion of a single adult colonic stem cell.