Protocol for generating reproducible miniaturized controlled midbrain organoids

Muwan Chen; Jonathan Christos Niclis; Mark Denham

doi:10.1016/j.xpro.2023.102451

Protocol for generating reproducible miniaturized controlled midbrain organoids

Muwan Chen^*, Jonathan Christos Niclis, Mark Denham^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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.

Original language	English
Article number	102451
Journal	STAR Protocols
Volume	4
Issue	3
DOIs	https://doi.org/10.1016/j.xpro.2023.102451
Publication status	Published - Sept 2023

Keywords

Neuroscience
Organoids

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10.1016/j.xpro.2023.102451Licence: CC BY

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title = "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 AggreWellTM400, 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.",

keywords = "Neuroscience, Organoids",

author = "Muwan Chen and Niclis, {Jonathan Christos} and Mark Denham",

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year = "2023",

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AU - Niclis, Jonathan Christos

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AB - 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 AggreWellTM400, 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.

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KW - Organoids

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ER -

Protocol for generating reproducible miniaturized controlled midbrain organoids

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Keywords

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