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Bioengineering

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

Article DOI: 10.3791/61143-v • 10:40 min • June 13th, 2020
June 13th, 2020 •
1,2, 1, 1, 1, 1,2,3, 4, 4, 4, 2, 1
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

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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.

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Scalable Generation Mature Cerebellar Organoids Human Pluripotent Stem Cells Immunostaining Reproducible Chemical Defined Conditions Single Use Bioreactors Cell Detachment Medium Human IPSC's Enzymatic Digestion Complete Cell Culture Medium Sterile Conical Tube Centrifuge Cell Pellet Culture Medium Hemocytometer Trypan Blue Dye Bioreactor Vessel Density Universal Base Unit Incubator Humidity Carbon Dioxide
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