Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells


Classic embryological studies have successfully applied genetics and cell biology principles to understand embryonic development. However, it remains unresolved how mechanics, as an integral driver of development, is involved in controlling tissue-scale cell fate patterning. Here we report a micropatterned human pluripotent stem (hPS)-cell-based neuroectoderm developmental model, in which pre-patterned geometrical confinement induces emergent patterning of neuroepithelial and neural plate border cells, mimicking neuroectoderm regionalization during early neurulation in vivo. In this hPS-cell-based neuroectoderm patterning model, two tissue-scale morphogenetic signals—cell shape and cytoskeletal contractile force—instruct neuroepithelial/neural plate border patterning via BMP-SMAD signalling. We further show that ectopic mechanical activation and exogenous BMP signalling modulation are sufficient to perturb neuroepithelial/neural plate border patterning. This study provides a useful microengineered, hPS-cell-based model with which to understand the biomechanical principles that guide neuroectoderm patterning and hence to study neural development and disease.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: Self-organized neuroectoderm patterning in circular hPS cell colonies.
Fig. 2: Self-organization of morphogenetic factors controls neuroectoderm patterning.
Fig. 3: Mechanics-guided neuroectoderm patterning is mediated by BMP-SMAD signalling.
Fig. 4: Mechanical force is sufficient for activating BMP-SMAD signalling and inducing NPB cell differentiation.
Fig. 5: BMP-SMAD signalling is required for mechanics-guided neuroectoderm patterning.


  1. 1.

    Wozniak, M. A. & Chen, C. S. Mechanotransduction in development: a growing role for contractility. Nat. Rev. Mol. Cell Biol. 10, 34–43 (2009).

    Article  Google Scholar 

  2. 2.

    Keller, R. Physical biology returns to morphogenesis. Science 338, 201–203 (2012).

    Article  Google Scholar 

  3. 3.

    Heller, E. & Fuchs, E. Tissue patterning and cellular mechanics. J. Cell Biol. 211, 219–231 (2015).

    Article  Google Scholar 

  4. 4.

    Chan, C. J., Heisenberg, C.-P. & Hiiragi, T. Coordination of morphogenesis and cell-fate specification in development. Curr. Biol. 27, R1024–R1035 (2017).

    Article  Google Scholar 

  5. 5.

    Gilmour, D., Rembold, M. & Leptin, M. From morphogen to morphogenesis and back. Nature 541, 311–320 (2017).

    Article  Google Scholar 

  6. 6.

    Schroeder, T. E. Neurulation in Xenopus laevis. An analysis and model based upon light and electron microscopy. J. Embryol. Exp. Morphol. 23, 427–462 (1970).

    Google Scholar 

  7. 7.

    Colas, J.-F. & Schoenwolf, G. C. Towards a cellular and molecular understanding of neurulation. Dev. Dynam. 221, 117–145 (2001).

    Article  Google Scholar 

  8. 8.

    Munoz-Sanjuan, I. & Brivanlou, A. H. Neural induction, the default model and embryonic stem cells. Nat. Rev. Neurosci. 3, 271–280 (2002).

    Article  Google Scholar 

  9. 9.

    Stern, C. D. Neural induction: old problem, new findings, yet more questions. Development 132, 2007–2021 (2005).

    Article  Google Scholar 

  10. 10.

    Bier, E. & De Robertis, E. M. BMP gradients: a paradigm for morphogen-mediated developmental patterning. Science 348, aaa5838 (2015).

  11. 11.

    Vijayraghavan, D. S. & Davidson, L. A. Mechanics of neurulation: From classical to current perspectives on the physical mechanics that shape, fold, and form the neural tube. Birth Defects Res. 109, 153–168 (2017).

    Article  Google Scholar 

  12. 12.

    Tesar, P. J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196–199 (2007).

    Article  Google Scholar 

  13. 13.

    O’Leary, T. et al. Tracking the progression of the human inner cell mass during embryonic stem cell derivation. Nat. Biotechnol. 30, 278–282 (2012).

    Article  Google Scholar 

  14. 14.

    Nakano, T. et al. Self-formation of optic cups and storable stratified neural retina from human ESCs. Cell Stem Cell 10, 771–785 (2012).

    Article  Google Scholar 

  15. 15.

    Lancaster, M. A. et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373–379 (2013).

    Article  Google Scholar 

  16. 16.

    Poh, Y.-C. et al. Generation of organized germ layers from a single mouse embryonic stem cell. Nat. Commun. 5, 4000 (2014).

    Article  Google Scholar 

  17. 17.

    Warmflash, A. et al. A method to recapitulate early embryonic spatial patterning in human embryonic stem cells. Nat. Methods 11, 847–854 (2014).

    Article  Google Scholar 

  18. 18.

    Ma, Z. et al. Self-organizing human cardiac microchambers mediated by geometric confinement. Nat. Commun. 6, 7413 (2015).

    Article  Google Scholar 

  19. 19.

    Shao, Y. et al. Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche. Nat. Mater. 16, 419–425 (2017).

    Article  Google Scholar 

  20. 20.

    Shao, Y. et al. A pluripotent stem cell-based model for post-implantation human amniotic sac development. Nat. Commun. 8, 208 (2017).

    Article  Google Scholar 

  21. 21.

    Tewary, M. et al. A stepwise model of reaction-diffusion and positional-information governs self-organized human peri-gastrulation-like patterning. Development 144, 4298–4312 (2017).

    Article  Google Scholar 

  22. 22.

    Chambers, S. M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of Smad signaling. Nat. Biotechnol. 27, 275–280 (2009).

    Article  Google Scholar 

  23. 23.

    Mica, Y. et al. Modeling neural crest induction, melanocyte specification, and disease-related pigmentation defects in hESCs and patient-specific iPSCs. Cell Rep. 3, 1140–1152 (2013).

    Article  Google Scholar 

  24. 24.

    Tchieu, J. et al. A modular platform for differentiation of human PSCs into all major ectodermal lineages. Cell Stem Cell 21, 399–410 (2017).

    Article  Google Scholar 

  25. 25.

    Hong, C. S. & Saint-Jeannet, J. P. The activity of Pax3 and Zic1 regulates three distinct cell fates at the neural plate border. Mol. Biol. Cell 18, 2192–2202 (2007).

    Article  Google Scholar 

  26. 26.

    Sun, Y. et al. Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells. Nat. Mater. 13, 599–604 (2014).

    Article  Google Scholar 

  27. 27.

    Sauka-Spengler, T. & Bronner-Fraser, M. A gene regulatory network orchestrates neural crest formation. Nat. Rev. Mol. Cell Biol. 9, 557–568 (2008).

    Article  Google Scholar 

  28. 28.

    Walther, C. & Gruss, P. PAX-6, a murine paired box gene, is expressed in the developing CNS. Development 113, 1435–1449 (1991).

    Google Scholar 

  29. 29.

    Milet, C. & Monsoro-Burq, A. H. Neural crest induction at the neural plate border in vertebrates. Dev. Biol. 366, 22–33 (2012).

    Article  Google Scholar 

  30. 30.

    Fu, J. et al. Mechanical regulation of cell function with geometrically modulated elastomeric substrates. Nat. Methods 7, 733–736 (2010).

    Article  Google Scholar 

  31. 31.

    Wilson, P. A. et al. Concentration-dependent patterning of the Xenopus ectoderm by BMP4 and its signal transducer Smad1. Development 124, 3177–3184 (1997).

    Google Scholar 

  32. 32.

    Michielin, F. et al. Microfluidic-assisted cyclic mechanical stimulation affects cellular membrane integrity in a human muscular dystrophy in vitro model. RSC Adv. 5, 98429–98439 (2015).

    Article  Google Scholar 

  33. 33.

    Xu, R.-H. et al. Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. Nat. Methods 2, 185–190 (2005).

    Article  Google Scholar 

  34. 34.

    Lippmann, E. S., Estevez-Silva, M. C. & Ashton, R. S. Defined human pluripotent stem cell culture enables highly efficient neuroepithelium derivation without small molecule inhibitors. Stem Cells 32, 1032–1042 (2014).

    Article  Google Scholar 

  35. 35.

    Etoc, F. et al. A balance between secreted inhibitors and edge sensing controls gastruloid self-organization. Dev. Cell 39, 302–315 (2016).

    Article  Google Scholar 

  36. 36.

    Nallet-Staub, F. et al. Cell density sensing alters TGF-β signaling in a cell-type-specific manner, independent from Hippo pathway activation. Dev. Cell 32, 640–651 (2015).

    Article  Google Scholar 

  37. 37.

    Varelas, X. et al. The Crumbs complex couples cell density sensing to Hippo-dependent control of the TGF-β-SMAD pathway. Dev. Cell 19, 831–844 (2010).

    Article  Google Scholar 

  38. 38.

    Dupont, S. et al. Role of YAP/TAZ in mechanotransduction. Nature 474, 179–183 (2011).

    Article  Google Scholar 

  39. 39.

    Wada, K.-I. et al. Hippo pathway regulation by cell morphology and stress fibers. Development 138, 3907–3914 (2011).

    Article  Google Scholar 

  40. 40.

    Sorrentino, G. et al. Metabolic control of YAP and TAZ by the mevalonate pathway. Nat. Cell Biol. 16, 357–366 (2014).

    Article  Google Scholar 

  41. 41.

    Kopf, J. et al. BMP growth factor signaling in a biomechanical context. BioFactors 40, 171–187 (2014).

    Article  Google Scholar 

  42. 42.

    de Croz‚, N., Maczkowiak, F. & Monsoro-Burq, A. H. Reiterative AP2a activity controls sequential steps in the neural crest gene regulatory network. Proc. Natl Acad. USA 108, 155–160 (2011).

    Article  Google Scholar 

  43. 43.

    Steventon, B. et al. Differential requirements of BMP and Wnt signalling during gastrulation and neurulation define two steps in neural crest induction. Development 136, 771–779 (2009).

    Article  Google Scholar 

  44. 44.

    Moury, J. D. & Schoenwolf, G. C. Cooperative model of epithelial shaping and bending during avian neurulation: autonomous movements of the neural plate, autonomous movements of the epidermis, and interactions in the neural plate/epidermis transition zone. Dev. Dynam. 204, 323–337 (1995).

    Article  Google Scholar 

  45. 45.

    Villa-Diaz, L. G. et al. Synthetic polymer coatings for long-term growth of human embryonic stem cells. Nat. Biotechnol. 28, 581–583 (2010).

    Article  Google Scholar 

  46. 46.

    Braam, S. R. et al. Recombinant vitronectin is a functionally defined substrate that supports human embryonic stem cell self-renewal via αVβ5 integrin. Stem Cells 26, 2257–2265 (2008).

    Article  Google Scholar 

  47. 47.

    Li, X. et al. Desktop aligner for fabrication of multilayer microfluidic devices. Rev. Sci. Instrum. 86, 075008 (2015).

    Article  Google Scholar 

  48. 48.

    Weng, S. et al. Mechanosensitive subcellular rheostasis drives emergent single-cell mechanical homeostasis. Nat. Mater. 15, 961–967 (2016).

    Article  Google Scholar 

Download references


We thank A. Liu for comments on the manuscript. This work is supported in part by the National Science Foundation (CMMI 1129611 and CBET 1149401 to J.F. and CMMI 1662835 to Y. Sun), the American Heart Association (12SDG12180025 to J.F.) and the Department of Mechanical Engineering at the University of Michigan. The Lurie Nanofabrication Facility at the University of Michigan, a member of the National Nanotechnology Infrastructure Network funded by the National Science Foundation, is acknowledged for support in microfabrication.

Author information




Y. Sun, X.X. and J.F. designed experiments; X.X. and Y. Sun performed differentiation assays; A.R.-I. and Y. Sun developed MATLAB scripts for image processing; X.X., Y. Sun, K.M.A.Y., Y.Z., S.W. and Y. Shao generated and analysed gene expression data; X.X. and Y.Y. conducted cell migration assays; L.S. provided Sox10-EGFP cells; Y. Sun, X.X., Y.C. and J.F. analysed data and wrote the manuscript. J.F. supervised the entire project. All authors edited and approved the manuscript.

Corresponding authors

Correspondence to Yubing Sun or Jianping Fu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Figures 1–20 and Supplementary Tables 1–3

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Xue, X., Sun, Y., Resto-Irizarry, A.M. et al. Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells. Nature Mater 17, 633–641 (2018).

Download citation

Further reading


Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing