Human brain wrinkling has been implicated in neurodevelopmental disorders and yet its origins remain unknown. Polymer gel models suggest that wrinkling emerges spontaneously due to compression forces arising during differential swelling, but these ideas have not been tested in a living system. Here, we report the appearance of surface wrinkles during the in vitro development and self-organization of human brain organoids in a microfabricated compartment that supports in situ imaging over a timescale of weeks. We observe the emergence of convolutions at a critical cell density and maximal nuclear strain, which are indicative of a mechanical instability. We identify two opposing forces contributing to differential growth: cytoskeletal contraction at the organoid core and cell-cycle-dependent nuclear expansion at the organoid perimeter. The wrinkling wavelength exhibits linear scaling with tissue thickness, consistent with balanced bending and stretching energies. Lissencephalic (smooth brain) organoids display reduced convolutions, modified scaling and a reduced elastic modulus. Although the mechanism here does not include the neuronal migration seen in vivo, it models the physics of the folding brain remarkably well. Our on-chip approach offers a means for studying the emergent properties of organoid development, with implications for the embryonic human brain.

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We are grateful for the help of S. Viukov, T. Levy and T. Sapir, and for fruitful discussions with S. Safran, A. Tayar and R. Bar-Ziv from the Weizmann Institute of Science. Device fabrication was carried out with assistance from A. Jahanfard, laser microdissection with assistance from Y. Fried, and RNA sequencing and analysis with the advice of H. Keren-Shaul, R. Kohen and T. Olender. Plasmid gifts were received from M. Davidson, Florida State University and J. LoTurco, University of Connecticut. MRI scans were provided by N. Bahi-Buisson, French Institute of Health and Medical Research. O.R. is the incumbent of the Bernstein-Mason Chair of Neurochemistry. E.K. is a Koshland fellow. The research has been supported by the Legacy Heritage Biomedical Program of the Israel Science Foundation (grant no. 2041/16), ERA-NET Neuron with support of the IMOH (grant no. 3-0000-12276), European Cooperation on Science and Technology (COST Action CA16118), Weizmann-FAPESP supported by a research grant from Sergio and Sonia Lozinsky, Nella and Leon Benoziyo Center for Neurological Diseases, Jeanne and Joseph Nissim Foundation for Life Sciences Research, Wohl Biology Endowment Fund, Lulu P. & David J. Levidow Fund for Alzheimers Diseases and Neuroscience Research, the Helen and Martin Kimmel Stem Cell Research Institute, the Kekst Family Institute for Medical Genetics, the David and Fela Shapell Family Center for Genetic Disorders Research. J.H.H. is a New York Stem Cell Foundation (NYSCF)–Robertson Investigator and is supported by research grants from the European Research Council (ERC-CoG2016 CellNaivety), Flight Attendant Medical Research Council (FAMRI), Israel Science Foundation Morasha Program, Nella and Leon Benoziyo Center for Neurological Diseases.

Author information


  1. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

    • Eyal Karzbrun
    • , Aditya Kshirsagar
    • , Jacob H. Hanna
    •  & Orly Reiner
  2. Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel

    • Sidney R. Cohen


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E.K., A.K. and O.R. planned and conducted experiments. S.C. designed and conducted AFM experiments. J.H. planned and assisted in hES-related experiments. The manuscript was prepared with inputs by all the authors.

Competing interests

J.H.H. is an advisor to Biological Industries Ltd and Accelta Ltd, and had issued patent applications and commercial licences related to certain human stem cell methods used herein.

Corresponding author

Correspondence to Orly Reiner.

Supplementary information

  1. Supplementary Movie 1

    Developing wild-type organoid. Fluorescence time-lapse images showing nuclear motion and cell division during cell-cycle. Green: Lifeact-GFP, RED: H2B-mCherry. Film duration 36 hours, time steps 10 minutes

  2. Supplementary Movie 2

    Developing mutant organoid. Fluorescence time-lapse images showing nuclear motion and cell division in LIS1+/– mutant organoids. Green: Lifeact-GFP, RED: H2B-mCherry. Film duration 5 hours, time steps 3 minutes

  3. Supplementary Information

    Supplementary Table S1, Supplementary Movie Captions S1 and S2, Supplementary Figures S1–S21

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