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

ABSTRACT: 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.

SUBMITTER: Takahashi J

PROVIDER: S-EPMC10320313 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature

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

Takahashi Junichi J Sugihara Hady Yuki HY Kato Shu S Nagata Sayaka S Okamoto Ryuichi R Mizutani Tomohiro T

STAR protocols 20230622 3

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 bioreac ...[more]

PMID: 37352105

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