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Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny

Author(s)
Yin, Xiaolei; Clevers, Hans; Farin, Henner F.; van Es, Johan H.; Karp, Jeffrey Michael; Langer, Robert S; ... Show more Show less
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Abstract
Although Lgr5[superscript +] 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[superscript +] 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[superscript +] 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.
Date issued
2013-12
URI
http://hdl.handle.net/1721.1/101203
Department
Harvard University--MIT Division of Health Sciences and Technology; Massachusetts Institute of Technology. Department of Chemical Engineering; Koch Institute for Integrative Cancer Research at MIT
Journal
Nature Methods
Publisher
Nature Publishing Group
Citation
Yin, Xiaolei, Henner F Farin, Johan H van Es, Hans Clevers, Robert Langer, and Jeffrey M Karp. “Niche-Independent High-Purity Cultures of Lgr5[superscript +] Intestinal Stem Cells and Their Progeny.” Nat Meth 11, no. 1 (December 1, 2013): 106–112.
Version: Author's final manuscript
ISSN
1548-7091
1548-7105

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