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Generation of cerebral organoids from human pluripotent stem cells

Nature Protocols volume 9, pages 2329–2340 (2014)Cite this article

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Abstract

Human brain development exhibits several unique aspects, such as increased complexity and expansion of neuronal output, that have proven difficult to study in model organisms. As a result, in vitro approaches to model human brain development and disease are an intense area of research. Here we describe a recently established protocol for generating 3D brain tissue, so-called cerebral organoids, which closely mimics the endogenous developmental program. This method can easily be implemented in a standard tissue culture room and can give rise to developing cerebral cortex, ventral telencephalon, choroid plexus and retinal identities, among others, within 1–2 months. This straightforward protocol can be applied to developmental studies, as well as to the study of a variety of human brain diseases. Furthermore, as organoids can be maintained for more than 1 year in long-term culture, they also have the potential to model later events such as neuronal maturation and survival.

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Figure 1: Schematic diagram of the cerebral organoid method and timing.
Figure 2: Progression of cerebral organoid development from human PSCs.
Figure 3: Examples of suitable and suboptimal organoids at various stages.
Figure 4: Staining for brain regions and neuronal cell identities.

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Acknowledgements

We are grateful to members of the Knoblich laboratory for their technical expertise and feedback, and particularly to M. Renner and A. Peer for their experimental support. We also thank the Stem Cell and BioOptics core facilities of IMBA and the Institute of Molecular Pathology for their technical support. M.A.L. received funding from a European Molecular Biology Organization (EMBO) postdoctoral fellowship, a Helen Hay Whitney postdoctoral fellowship and a Marie Curie International Incoming Fellowship. Work in J.A.K.'s laboratory is supported by the Austrian Academy of Sciences, the Austrian Science Fund (FWF; projects Z153-B09 and I552-B19) and an advanced grant from the European Research Council.

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Authors and Affiliations

  1. Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna, Austria

    Madeline A Lancaster & Juergen A Knoblich

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  1. Madeline A Lancaster
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  2. Juergen A Knoblich
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J.A.K. and M.A.L. wrote the manuscript, and M.A.L. performed the experiments.

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Correspondence to Madeline A Lancaster or Juergen A Knoblich.

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Lancaster, M., Knoblich, J. Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9, 2329–2340 (2014). https://doi.org/10.1038/nprot.2014.158

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