1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
12
Lancaster, M. A. et al. Cerebral organoids model human brain development
and microcephaly. Nature 501, 373–379 (2013).
Kadoshima, T. et al. Self-organization of axial polarity, inside-out layer
pattern, and species-specific progenitor dynamics in human ES cell-derived
neocortex. Proc. Natl Acad. Sci. USA 110, 20284–20289 (2013).
Di Lullo, E. & Kriegstein, A. R. The use of brain organoids to investigate
neural development and disease. Nat. Rev. Neurosci. 18, 573–584 (2017).
Pasca, S. P. The rise of three-dimensional human brain cultures. Nature 553,
437–445 (2018).
Trujillo, C. A. et al. Complex oscillatory waves emerging from cortical
organoids model early human brain network development. Cell Stem Cell 25,
558–569 (2019).
Quadrato, G. et al. Cell diversity and network dynamics in photosensitive
human brain organoids. Nature 545, 48–53 (2017).
Camp, J. G. et al. Human cerebral organoids recapitulate gene expression
programs of fetal neocortex development. Proc. Natl Acad. Sci. USA 112,
15672–15677 (2015).
Giandomenico, S. L. et al. Cerebral organoids at the air-liquid interface
generate diverse nerve tracts with functional output. Nat. Neurosci. 22,
669–679 (2019).
Muguruma, K., Nishiyama, A., Kawakami, H., Hashimoto, K. & Sasai, Y. Selforganization of polarized cerebellar tissue in 3D culture of human pluripotent
stem cells. Cell Rep. 10, 537–550 (2015).
Seto, Y. & Eiraku, M. Toward the formation of neural circuits in human brain
organoids. Curr. Opin. Cell Biol. 61, 86–91 (2019).
Takebe, T. & Wells, J. M. Organoids by design. Science 364, 956–959 (2019).
Dupont, S. et al. Role of YAP/TAZ in mechanotransduction. Nature 474,
179–183 (2011).
Sunyer, R. et al. Collective cell durotaxis emerges from long-range intercellular
force transmission. Science 353, 1157–1161 (2016).
Nerurkar, N. L., Lee, C., Mahadevan, L. & Tabin, C. J. Molecular control of
macroscopic forces drives formation of the vertebrate hindgut. Nature 565,
480–484 (2019).
Ito, Y. et al. Turbulence activates platelet biogenesis to enable clinical scale
ex vivo production. Cell 174, 636–648.e618 (2018).
Dahl-Jensen, S. & Grapin-Botton, A. The physics of organoids: a biophysical
approach to understanding organogenesis. Development 144, 946–951 (2017).
Goto-Silva, L. et al. Computational fluid dynamic analysis of physical forces
playing a role in brain organoid cultures in two different multiplex platforms.
BMC Dev. Biol. 19, 3 (2019).
Montes-Olivas, S., Marucci, L. & Homer, M. Mathematical models of
organoid cultures. Front. Genet. 10, 873 (2019).
Pala, R., Alomari, N. & Nauli, S. M. Primary cilium-dependent
signaling mechanisms. Int. J. Mol. Sci. 18, https://doi.org/10.3390/
ijms18112272 (2017).
Elliott, K. H. & Brugmann, S. A. Sending mixed signals: ciliadependent signaling during development and disease. Dev. Biol. 447, 28–41
(2019).
21. Rallu, M. et al. Dorsoventral patterning is established in the telencephalon of
mutants lacking both Gli3 and Hedgehog signaling. Development 129,
4963–4974 (2002).
22. Gulacsi, A. & Anderson, S. A. Shh maintains Nkx2.1 in the MGE by a Gli3independent mechanism. Cereb. Cortex 16, i89–i95 (2006). Suppl 1.
23. Hasenpusch-Theil, K. & Theil, T. The multifaceted roles of primary cilia in the
development of the cerebral cortex. Front Cell Dev. Biol. 9, 630161 (2021).
24. Xavier, G. M. et al. Hedgehog receptor function during craniofacial
development. Dev. Biol. 415, 198–215 (2016).
25. Nechipurenko, I. V. et al. A conserved role for girdin in basal body positioning
and ciliogenesis. Dev. Cell 38, 493–506 (2016).
26. Matsumoto, M. et al. Dynamic changes in ultrastructure of the primary cilium
in migrating neuroblasts in the postnatal brain. J. Neurosci. 39, 9967–9988
(2019).
27. Wilsch-Bräuninger, M., Peters, J., Paridaen, J. T. & Huttner, W. B. Basolateral
rather than apical primary cilia on neuroepithelial cells committed to
delamination. Development 139, 95–105 (2012).
28. Li, Y. et al. Implications of GABAergic neurotransmission in Alzheimer’s
disease. Front Aging Neurosci. 8, 31 (2016).
29. Kondo, T. et al. Modeling Alzheimer’s disease with iPSCs reveals stress
phenotypes associated with intracellular Abeta and differential drug
responsiveness. Cell Stem Cell 12, 487–496 (2013).
30. Nasu, M. et al. Robust formation and maintenance of continuous stratified
cortical neuroepithelium by laminin-containing matrix in mouse ES cell
culture. PLoS ONE 7, e53024 (2012).
31. Higginbotham, H. et al. Arl13b-regulated cilia activities are essential for
polarized radial glial scaffold formation. Nat. Neurosci. 16, 1000–1007 (2013).
32. Nampe, D., Joshi, R., Keller, K., Zur Nieden, N. I. & Tsutsui, H. Impact of
fluidic agitation on human pluripotent stem cells in stirred suspension culture.
Biotechnol. Bioeng. 114, 2109–2120 (2017).
33. Mercurio, S., Serra, L. & Nicolis, S. K. More than just stem cells: functional
roles of the transcription factor Sox2 in differentiated glia and neurons. Int. J.
Mol. Sci. 20, https://doi.org/10.3390/ijms20184540 (2019).
34. Cavallaro, M. et al. Impaired generation of mature neurons by neural stem
cells from hypomorphic Sox2 mutants. Development 135, 541–557 (2008).
35. Higginbotham, H. et al. Arl13b in primary cilia regulates the migration and
placement of interneurons in the developing cerebral cortex. Dev. Cell 23,
925–938 (2012).
36. Baudoin, J. P. et al. Tangentially migrating neurons assemble a primary cilium that
promotes their reorientation to the cortical plate. Neuron 76, 1108–1122 (2012).
37. Qian, X. et al. Brain-region-specific organoids using mini-bioreactors for
modeling ZIKV exposure. Cell 165, 1238–1254 (2016).
38. Sequerra, E. B., Miyakoshi, L. M., Fróes, M. M., Menezes, J. R. & HedinPereira, C. Generation of glutamatergic neurons from postnatal and adult
subventricular zone with pyramidal-like morphology. Cereb. Cortex 20,
2583–2591 (2010).
39. Lai, T. et al. SOX5 controls the sequential generation of distinct corticofugal
neuron subtypes. Neuron 57, 232–247 (2008).
40. Nikouei, K., Muñoz-Manchado, A. B. & Hjerling-Leffler, J. BCL11B/CTIP2 is
highly expressed in GABAergic interneurons of the mouse somatosensory
cortex. J. Chem. Neuroanat. 71, 1–5 (2016).
41. Takahashi, K. et al. Induction of pluripotent stem cells from adult human
fibroblasts by defined factors. Cell 131, 861–872 (2007).
42. Sutcliffe, M. & Lancaster, M. A. A simple method of generating 3D brain
organoids using standard laboratory equipment. Methods Mol. Biol. 1576,
1–12 (2019).
43. Lambert, M. P. et al. Monoclonal antibodies that target pathological
assemblies of Abeta. J. Neurochem. 100, 23–35 (2007).
Acknowledgements
We would like to express our sincere gratitude to all of our co-workers and collaborators:
Kayoko Tsukita, Takako Enami, and Ayako Nagahashi for their technical support, Mikie
Iijima, Nozomi Kawabata, Tomomi Urai, and Miho Nagata for their valuable administrative support, and Minako Tateno for critical reading of the manuscript. This research
was funded in part by a grant for Core Center for iPS Cell Research of the Research
Center Network for Realization of Regenerative Medicine from AMED (H.I.) and the
Uehara Memorial Foundation (H.I.) and partially supported by the Center of Innovation
Program from MEXT and JST (H.I.).
Author contributions
H.I. conceived the project. DN.AS., K.I., I.I., R.K., S.S., T.O., Y.K., T.K., Y.Y. and A.W.
performed the experiments. W.L.K. provided antibodies. DN.AS., K.I., T.K. and H.I.
wrote the manuscript.
Competing interests
The authors declare no competing interests.
COMMUNICATIONS BIOLOGY | (2021)4:1213 | https://doi.org/10.1038/s42003-021-02719-5 | www.nature.com/commsbio
A Self-archived copy in
Kyoto University Research Information Repository
https://repository.kulib.kyoto-u.ac.jp
COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02719-5
Additional information
Supplementary information The online version contains supplementary material
available at https://doi.org/10.1038/s42003-021-02719-5.
Correspondence and requests for materials should be addressed to Haruhisa Inoue.
Peer review information Communications Biology thanks the anonymous reviewers for
their contribution to the peer review of this work. Primary Handling Editor: Christina
Karlsson Rosenthal. Peer reviewer reports are available.
Reprints and permission information is available at http://www.nature.com/reprints
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
ARTICLE
Open Access This article is licensed under a Creative Commons
Attribution 4.0 International License, which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative
Commons license, and indicate if changes were made. The images or other third party
material in this article are included in the article’s Creative Commons license, unless
indicated otherwise in a credit line to the material. If material is not included in the
article’s Creative Commons license and your intended use is not permitted by statutory
regulation or exceeds the permitted use, you will need to obtain permission directly from
the copyright holder. To view a copy of this license, visit http://creativecommons.org/
licenses/by/4.0/.
© The Author(s) 2021
COMMUNICATIONS BIOLOGY | (2021)4:1213 | https://doi.org/10.1038/s42003-021-02719-5 | www.nature.com/commsbio
13
...