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Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny.


ABSTRACT: Although Lgr5(+) intestinal stem cells have been expanded in vitro as organoids, homogeneous culture of these cells has not been possible thus far. Here we show that two small molecules, CHIR99021 and valproic acid, synergistically maintain self-renewal of mouse Lgr5(+) intestinal stem cells, resulting in nearly homogeneous cultures. The colony-forming efficiency of cells from these cultures is ~100-fold greater than that of cells cultured in the absence of CHIR99021 and valproic acid, and multilineage differentiation ability is preserved. We made use of these homogeneous cultures to identify conditions employing simultaneous modulation of Wnt and Notch signaling to direct lineage differentiation into mature enterocytes, goblet cells and Paneth cells. Expansion in these culture conditions may be feasible for Lgr5(+) cells from the mouse stomach and colon and from the human small intestine. These methods provide new tools for the study and application of multiple intestinal epithelial cell types.

SUBMITTER: Yin X 

PROVIDER: S-EPMC3951815 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny.

Yin Xiaolei X   Farin Henner F HF   van Es Johan H JH   Clevers Hans H   Langer Robert R   Karp Jeffrey M JM  

Nature methods 20131201 1


Although Lgr5(+) intestinal stem cells have been expanded in vitro as organoids, homogeneous culture of these cells has not been possible thus far. Here we show that two small molecules, CHIR99021 and valproic acid, synergistically maintain self-renewal of mouse Lgr5(+) intestinal stem cells, resulting in nearly homogeneous cultures. The colony-forming efficiency of cells from these cultures is ~100-fold greater than that of cells cultured in the absence of CHIR99021 and valproic acid, and multi  ...[more]

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