Protocol for generating reproducible miniaturized controlled midbrain organoids

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(TM)400, and maintaini...

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Detalles Bibliográficos
Autores principales: Chen, Muwan, Niclis, Jonathan Christos, Denham, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10382973/
https://www.ncbi.nlm.nih.gov/pubmed/37481727
http://dx.doi.org/10.1016/j.xpro.2023.102451
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author Chen, Muwan
Niclis, Jonathan Christos
Denham, Mark
author_facet Chen, Muwan
Niclis, Jonathan Christos
Denham, Mark
author_sort Chen, Muwan
collection PubMed
description 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(TM)400, and maintaining them in an EB-Disk(360)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 analysis. This protocol is suitable for compound screening and disease modeling studies.
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spelling pubmed-103829732023-07-30 Protocol for generating reproducible miniaturized controlled midbrain organoids Chen, Muwan Niclis, Jonathan Christos Denham, Mark STAR Protoc Protocol 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(TM)400, and maintaining them in an EB-Disk(360)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 analysis. This protocol is suitable for compound screening and disease modeling studies. Elsevier 2023-07-21 /pmc/articles/PMC10382973/ /pubmed/37481727 http://dx.doi.org/10.1016/j.xpro.2023.102451 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Chen, Muwan
Niclis, Jonathan Christos
Denham, Mark
Protocol for generating reproducible miniaturized controlled midbrain organoids
title Protocol for generating reproducible miniaturized controlled midbrain organoids
title_full Protocol for generating reproducible miniaturized controlled midbrain organoids
title_fullStr Protocol for generating reproducible miniaturized controlled midbrain organoids
title_full_unstemmed Protocol for generating reproducible miniaturized controlled midbrain organoids
title_short Protocol for generating reproducible miniaturized controlled midbrain organoids
title_sort protocol for generating reproducible miniaturized controlled midbrain organoids
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10382973/
https://www.ncbi.nlm.nih.gov/pubmed/37481727
http://dx.doi.org/10.1016/j.xpro.2023.102451
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