Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
STAR Protoc. 2023 Sep 15;4(3):102374. doi: 10.1016/j.xpro.2023.102374. Epub 2023 Jun 22.
Current techniques for producing induced-pluripotent-stem-cell-derived mid/hindgut spheroids have faced major hurdles in consistency and reproducibility. Here, we present a protocol that uses mid/hindgut cells to generate homogeneous spheroids that subsequently mature into human intestinal organoids (HIOs). We describe steps for stepwise differentiation and spheroid formation using a 96-well plate. We then detail cell maturation in a suspended state and the implementation of a rotational bioreactor platform to maximize the culture efficiency of larger HIOs. For complete details on the use and execution of this protocol, please refer to Takahashi et al..
目前，生成诱导多能干细胞衍生的中肠/后肠类器官球体的技术在一致性和可重复性方面面临着重大的障碍。在这里，我们提出了一种使用中肠/后肠细胞来生成同质球体的方案，这些球体随后成熟为人类肠道类器官（HIOs）。我们描述了使用 96 孔板逐步分化和形成球体的步骤。然后，我们详细介绍了在悬浮状态下使细胞成熟的过程，并实施了旋转生物反应器平台，以最大化更大的 HIO 培养效率。有关此方案的使用和执行的完整详细信息，请参阅 Takahashi 等人的研究。
