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A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells
Hajime Uchida, … , Akihiro Umezawa, Hidenori Akutsu
Hajime Uchida, … , Akihiro Umezawa, Hidenori Akutsu
Published January 12, 2017
Citation Information: JCI Insight. 2017;2(1):e86492. https://doi.org/10.1172/jci.insight.86492.
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Categories: Technical Advance Development Stem cells

A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells

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Abstract

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.

Authors

Hajime Uchida, Masakazu Machida, Takumi Miura, Tomoyuki Kawasaki, Takuya Okazaki, Kengo Sasaki, Seisuke Sakamoto, Noriaki Ohuchi, Mureo Kasahara, Akihiro Umezawa, Hidenori Akutsu

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Figure 1

Generation of peristaltic gut organoids from human pluripotent stem cells on a patterning substrate.

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Generation of peristaltic gut organoids from human pluripotent stem cell...
(A) Illustration of gut organoid generation from human pluripotent stem cells cultured on a single device. (B) A micropatterned substrate for cell growth was produced by coating a small mesh net, 1,500 μm in diameter (left panel), with a bioactive polymer (scale bar: 15 mm). The right panel shows that human embryonic stem cells attached and grew only within each zone (scale bar: 500 μm). (C) Time course of organoid growth in culture. The organoid at day 63 is surrounded by intestinal columnar epithelium layers adjacent to mesenchymal tissues; these epithelial layers are delineated by the dotted white line (high-magnification view of the area within the red box). Scale bar: 200 μm (top row and right bottom); 50 μm (left bottom). (D) Six organoids that exhibited gut-like motor activity were maintained in culture at day 129 and did not coalesce. Scale bar: 5 mm.
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