A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells
A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells
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DOI：
10.1172/jci.insight.86492
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发表时间：
2017-01-12
期刊：
影响因子：
8
通讯作者：
Akutsu, Hidenori
中科院分区：
文献类型：
--
作者：
Uchida, Hajime;Machida, Masakazu;Akutsu, Hidenori
功能性肠由来自所有3个初级胚层的细胞类型组成，并通过高度协调和连续的发育过程产生。人类多能干细胞（hPSC）的定向分化已被证明可以产生肠道特异性细胞类型;然而，这些结构不会再现不同胚层细胞类型之间的关键功能相互作用。在这里，我们开发了一种简单的方案，用于在无异种条件下从hPSC产生成熟的功能性肠类器官。发现此处产生的干细胞衍生的肠道类器官含有不同类型的肠细胞，包括肠上皮细胞、杯状细胞、潘氏细胞和肠内分泌细胞，这些细胞来源于所有3个胚层;此外，它们表现出肠道功能，包括肽吸收，并显示出对组胺和抗胆碱能药物刺激的神经支配的肠运动。重要的是，使用这种无异种系统获得的肠道类器官可以在培养物中稳定地维持很长时间，并成功地在体内移植。我们用于从hPSC产生肠道类器官的无异种方法为研究人类肠道疾病和药理学测试提供了平台。
Functional intestines are composed of cell types from all 3 primary germ layers and are generated through a highly orchestrated and serial developmental process. Directed differentiation of human pluripotent stem cells (hPSCs) has been shown to yield gut-specific cell types; however, these structures do not reproduce critical functional interactions between cell types of different germ layers. Here, we developed a simple protocol for the generation of mature functional intestinal organoids from hPSCs under xenogeneic-free conditions. The stem cell-derived gut organoids produced here were found to contain distinct types of intestinal cells, including enterocytes, goblet cells, Paneth cells, and enteroendocrine cells, that were derived from all 3 germ layers; moreover, they demonstrated intestinal functions, including peptide absorption, and showed innervated bowel movements in response to stimulation with histamine and anticholinergic drugs. Importantly, the gut organoids obtained using this xenogeneic-free system could be stably maintained in culture for prolonged periods and were successfully engrafted in vivo. Our xenogeneic-free approach for generating gut organoids from hPSCs provides a platform for studying human intestinal diseases and for pharmacological testing.