Murine Colonic Organoid Culture System and Downstream Assay Applications
Colonic organoids, three-dimensional colonic crypts grown in vitro that show realistic microanatomy, have many potential applications for studying physiology, developmental biology, and pathophysiology of intestinal diseases including inflammatory bowel disease and colorectal cancer. Here, we describe detailed protocols for mouse colonic crypt isolation, organoid culture, and downstream applications. Specific culture strategies including growth factor enriched Matrigel and Wnt and R-spondin conditioned media serve as key factors for enhancing the growth and cost efficiency of colonic organoid cultures.
Keywords:Niche-independent crypt expansion Stem cells In vitro primary culture
This work was supported by funding from NIH R35CA197707 and P30ES023512. We would like to thank Drs. Hans Clevers, Nick Barker, and Toshiro Sato for initial guidance in organoid culture technique.
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