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Protocol for generating reproducible miniaturized controlled midbrain organoids.

ABSTRACT: Here, we present a protocol for generating miniaturized controlled midbrain organoids (MiCOs) of reproducible size and cellular composition, without a necrotic center. We describe steps for maintaining and passaging human pluripotent stem cells, generating MiCOs using AggreWell^TM400, and maintaining them in an EB-Disk³⁶⁰on an orbital shaker, eliminating the need for Matrigel or a spinner flask and preventing organoid fusion. We then detail organoid collection for different endpoint analysis. This protocol is suitable for compound screening and disease modeling studies.

SUBMITTER: Chen M

PROVIDER: S-EPMC10382973 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Protocol for generating reproducible miniaturized controlled midbrain organoids.

Chen Muwan M Niclis Jonathan Christos JC Denham Mark M

STAR protocols 20230721 3

Here, we present a protocol for generating miniaturized controlled midbrain organoids (MiCOs) of reproducible size and cellular composition, without a necrotic center. We describe steps for maintaining and passaging human pluripotent stem cells, generating MiCOs using AggreWell™400, and maintaining them in an EB-Disk<sup>360</sup>on an orbital shaker, eliminating the need for Matrigel or a spinner flask and preventing organoid fusion. We then detail organoid collection for different endpoint ana ...[more]

PMID: 37481727

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