Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny
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 +] intes...
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2016
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| Online Access: | http://hdl.handle.net/1721.1/101203 https://orcid.org/0000-0001-8624-8928 https://orcid.org/0000-0003-4255-0492 |
| _version_ | 1826200851715194880 |
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| author | Yin, Xiaolei Clevers, Hans Farin, Henner F. van Es, Johan H. Karp, Jeffrey Michael Langer, Robert S |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Yin, Xiaolei Clevers, Hans Farin, Henner F. van Es, Johan H. Karp, Jeffrey Michael Langer, Robert S |
| author_sort | Yin, Xiaolei |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:42:44Z |
| format | Article |
| id | mit-1721.1/101203 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:42:44Z |
| publishDate | 2016 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1012032022-09-27T21:25:33Z Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny Yin, Xiaolei Clevers, Hans Farin, Henner F. van Es, Johan H. Karp, Jeffrey Michael Langer, Robert S 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 Yin, Xiaolei Langer, Robert Karp, Jeffrey Michael 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. National Institutes of Health (U.S.) (Grant DE013023) Harvard Institute of Translational Immunology/Helmsley Trust Pilot Grant in Crohn's Disease 2016-02-17T21:26:43Z 2016-02-17T21:26:43Z 2013-12 2013-07 Article http://purl.org/eprint/type/JournalArticle 1548-7091 1548-7105 http://hdl.handle.net/1721.1/101203 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. https://orcid.org/0000-0001-8624-8928 https://orcid.org/0000-0003-4255-0492 en_US http://dx.doi.org/10.1038/nmeth.2737 Nature Methods Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Nature Publishing Group PMC |
| spellingShingle | Yin, Xiaolei Clevers, Hans Farin, Henner F. van Es, Johan H. Karp, Jeffrey Michael Langer, Robert S Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title | Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title_full | Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title_fullStr | Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title_full_unstemmed | Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title_short | Niche-independent high-purity cultures of Lgr5[superscript +] intestinal stem cells and their progeny |
| title_sort | niche independent high purity cultures of lgr5 superscript intestinal stem cells and their progeny |
| url | http://hdl.handle.net/1721.1/101203 https://orcid.org/0000-0001-8624-8928 https://orcid.org/0000-0003-4255-0492 |
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