STAR Protocols (Sep 2023)

Protocol for generating reproducible miniaturized controlled midbrain organoids

  • Muwan Chen,
  • Jonathan Christos Niclis,
  • Mark Denham

Journal volume & issue
Vol. 4, no. 3
p. 102451

Abstract

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Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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