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Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining
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Bioengineering
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JoVE Journal Bioengineering
Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining

Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining

DOI:
10.3791/61143-v

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10:40 min

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June 13, 2020

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, , , , , , , , ,
1iBB – Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior Técnico,Universidade de Lisboa, 2Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina,Universidade de Lisboa, 3The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus,Universidade de Lisboa, 4PBS Biotech, Inc, Camarillo, CA, USA

Chapters

  • 00:04Introduction
  • 00:23Seeding the Bioreactor
  • 02:06Differentiation and Maturation of Aggregates
  • 03:26Preparing Organoids for Cryosectioning
  • 05:47Immunostaining
  • 08:27Results: Efficient Cerebellar Differentiation and Maturation
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a dynamic culture system to produce controlled size aggregates of human pluripotent stem cells and further stimulate differentiation in cerebellar organoids under chemically-defined and feeder-free conditions using a single-use bioreactor.

Tags

Keywords: Cerebellar OrganoidsHuman Pluripotent Stem CellsScalable GenerationSingle-use BioreactorsImmunostainingIPSC AggregatesGfCDM Differentiation MediumCell DetachmentCentrifugationCell CountingBioreactor CultivationMedium ExchangeOrganoid Storage

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