Takahashi Junichi, Sugihara Hady Yuki, Kato Shu, Kawasaki Sho, Nagata Sayaka, Okamoto Ryuichi, Mizutani Tomohiro
Department of Gastroenterology and Hepatology, Institute of Science Tokyo, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
Department of Gastroenterology and Hepatology, Institute of Science Tokyo, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
Cell Rep Methods. 2024 Dec 16;4(12):100930. doi: 10.1016/j.crmeth.2024.100930. Epub 2024 Dec 10.
Human intestinal organoids (HIOs) derived from human pluripotent stem cells (hPSCs) are promising resources for intestinal regenerative therapy as they recapitulate both endodermal and mesodermal components of the intestine. However, due to their hPSC-line-dependent mesenchymal development and spherical morphology, HIOs have limited applicability beyond basic research and development. Here, we demonstrate the incorporation of separately differentiated mesodermal and mid/hindgut cells into assembled spheroids to stabilize mesenchymal growth in HIOs. In parallel, we generate tubular intestinal constructs (assembled human intestinal tubules [a-HITs]) by leveraging the high aggregative property of assembled spheroids. Through rotational culture in a bioreactor, a-HITs self-organize to develop epithelium and supportive mesenchyme. Upon mesenteric transplantation, a-HITs mature into centimeter-scale tubular intestinal tissue with complex architectures. Our aggregation- and suspension-based approach offers basic technology for engineering tubular intestinal tissue from hPSCs, which could be ultimately applied to the generation of the human intestine for clinical application.
源自人多能干细胞（hPSC）的人肠道类器官（HIO）是肠道再生治疗的有前景的资源，因为它们概括了肠道的内胚层和中胚层成分。然而，由于其依赖hPSC系的间充质发育和球形形态，HIO在基础研究和开发之外的适用性有限。在这里，我们展示了将分别分化的中胚层细胞和中/后肠细胞整合到组装的球体中，以稳定HIO中间充质的生长。同时，我们利用组装球体的高聚集特性生成管状肠道构建体（组装的人肠道小管[a-HIT]）。通过在生物反应器中进行旋转培养，a-HIT自我组织以发育上皮和支持性间充质。经肠系膜移植后，a-HIT成熟为具有复杂结构的厘米级管状肠道组织。我们基于聚集和悬浮的方法为从hPSC工程化管状肠道组织提供了基础技术，最终可应用于临床应用的人类肠道的生成。
