Protocol to generate large human intestinal organoids using a rotating bioreactor.

Autor:	Takahashi J; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. Electronic address: jtakahashi.gast@tmd.ac.jp., Sugihara HY; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan., Kato S; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan., Nagata S; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan., Okamoto R; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan., Mizutani T; Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. Electronic address: tmizutani.gast@tmd.ac.jp.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2023 Sep 15; Vol. 4 (3), pp. 102374. Date of Electronic Publication: 2023 Jun 22.
DOI:	10.1016/j.xpro.2023.102374
Abstrakt:	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. 1 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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