Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny.
Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny.
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DOI：
10.1038/nmeth.2737
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发表时间：
2014-01
期刊：
影响因子：
48
通讯作者：
Karp, Jeffrey M.
中科院分区：
文献类型：
--
作者：
Yin, Xiaolei;Farin, Henner F.;van Es, Johan H.;Clevers, Hans;Langer, Robert;Karp, Jeffrey M.
尽管Lgr5+肠干细胞已在体外扩增为类器官，但迄今为止还不可能对这些细胞进行同质培养。在这里，我们展示了两种小分子 CHIR99021 和丙戊酸协同维持小鼠 Lgr5+ 肠道干细胞的自我更新，从而产生几乎均质的培养物。来自这些培养物的细胞的集落形成效率比在不存在 CHIR99021 和丙戊酸的情况下培养的细胞高约 100 倍，并且保留了多谱系分化能力。我们利用这些均质培养物来鉴定同时调节 Wnt 和 Notch 信号传导以指导谱系分化为成熟肠上皮细胞、杯状细胞和潘氏细胞的条件。对于来自小鼠胃和结肠以及来自人类小肠的 Lgr5+ 细胞来说，在这些培养条件下进行扩增可能是可行的。这些方法为多种肠上皮细胞类型的研究和应用提供了新工具。
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.
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影响因子：
29.4
作者：
Farin, Henner F.;Van Es, Johan H.;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
4.8
作者：
Kim, Tae-Hee;Shivdasani, Ramesh A.
通讯作者：
Shivdasani, Ramesh A.
影响因子：
4.4
作者：
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通讯作者：
Honjo, T
影响因子：
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影响因子：
64.5
作者：
Snippert, Hugo J.;van der Flier, Laurens G.;Clevers, Hans
通讯作者：
Clevers, Hans