EX VIVO CULTURE, PROLIFERATION AND EXPANSION OF INTESTINAL EPITHELIUM

摘要

Methods are provided for long term culture of mammalian intestinal cells. Cultures are initiated with fragments of mammalian intestinal tissue, which are then maintained embedded in a gel substrate that provides an air-liquid interface. Intestinal epithelium in cultures of the invention can be continuously grown for extended periods of time. Mammalian intestinal cells cultured by the methods of the invention recapitulate features of intestinal growth in vivo.

主权项

1. A method for the long term culture of mammalian intestinal cells, wherein said culture provides for multilineage differentiation that maintains (a) the three-dimensional structure of intestinal epithelium and (b) differentiation markers characteristic of intestinal absorptive enterocytes, goblet cells and enteroendocrine cells, in which intestinal cells grow, differentiate and are viable for more than 3 months, said culture comprising a primary intestinal tissue explant cultured in the presence of exogenous R-spondin;
wherein the cells in the in vitro culture exhibit normal Wnt-signaling and are responsive to proteins that stimulate or inhibit intestinal growth.