Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by ImmunostainingTeresa P. Silva 1,2, Tiago G. Fernandes 1, Diogo E. S. Nogueira 1, Carlos A. V. Rodrigues 1, Evguenia P. Bekman 1,2,3, Yas Hashimura 4, Sunghoon Jung 4, Brian Lee 4, Maria Carmo-Fonseca 2, Joaquim M. S. Cabral 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
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.
