The Three-Dimensional Culture of Epithelial Organoids Derived from Embryonic Chicken Intestine
The intestinal epithelium isolated from chicken embryos in last 3 days of development can be used to establish the 3D culture of intestinal organoids. When fragments of epithelial tissue released by incubation with EGTA (2.5 mM, 2 h) are embedded in Matrigel matrix on cell culture inserts the formation of empty spheres covered by epithelial cells is observed in first 24 h of culture. The growth and survival of organoids are supported by the addition of R-spondin 1, Noggin, and prostaglandin E2 to the culture medium. The organoids are accompanied by myofibroblasts which become visible in the next 2 days of culture. The intestinal enteroids (free of myofibroblasts) can be obtained from adult chicken intestine.
Keywords:Intestinal epithelium Three-dimensional cell culture Epithelial organoids Chicken embryo Intestinal myofibroblasts
The authors acknowledge the financial support of Polish National Science Center (grant no. 2013/09/B/NZ9/00285 to M.P.)
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