Article | Published:

Self-organizing optic-cup morphogenesis in three-dimensional culture

Nature volume 472, pages 5156 (07 April 2011) | Download Citation

Abstract

Balanced organogenesis requires the orchestration of multiple cellular interactions to create the collective cell behaviours that progressively shape developing tissues. It is currently unclear how individual, localized parts are able to coordinate with each other to develop a whole organ shape. Here we report the dynamic, autonomous formation of the optic cup (retinal primordium) structure from a three-dimensional culture of mouse embryonic stem cell aggregates. Embryonic-stem-cell-derived retinal epithelium spontaneously formed hemispherical epithelial vesicles that became patterned along their proximal–distal axis. Whereas the proximal portion differentiated into mechanically rigid pigment epithelium, the flexible distal portion progressively folded inward to form a shape reminiscent of the embryonic optic cup, exhibited interkinetic nuclear migration and generated stratified neural retinal tissue, as seen in vivo. We demonstrate that optic-cup morphogenesis in this simple cell culture depends on an intrinsic self-organizing program involving stepwise and domain-specific regulation of local epithelial properties.

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Acknowledgements

We are grateful to S. Nakanishi, S. Yonemura, R. Ladher, K. Muguruma, H. Inomata and M. Ohgushi for comments and to members of the Sasai laboratory for discussion. We also thank Olympus, particularly Y. Saito, M. Suzuki and Y. Imai, for their help and discussion regarding the design, assembly and optimized utility of the incubator-combined multi-photon and confocal optic systems, and T. Sugitate and N. Saito at JPK Instruments for technical advice on the AFM assay. This work was supported by grants-in-aid from MEXT (Y.S., M.E., T.A.), the Knowledge Cluster Initiative at Kobe and the S-Innovation Project (Y.S., K.S.), and the Leading Project for Realization of Regenerative Medicine (Y.S).

Author information

Affiliations

  1. Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Mototsugu Eiraku
    • , Nozomu Takata
    • , Masako Kawada
    • , Eriko Sakakura
    •  & Yoshiki Sasai
  2. Four-Dimensional Tissue Analysis Unit, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Mototsugu Eiraku
    • , Eriko Sakakura
    •  & Yoshiki Sasai
  3. Department of Biomechanics, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan

    • Hiroki Ishibashi
    • , Satoru Okuda
    •  & Taiji Adachi
  4. Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita 565-0871, Japan

    • Kiyotoshi Sekiguchi
  5. Computational Cell Biomechanics Team, VCAD System Research Program, RIKEN, Wako 351-0198, Japan

    • Taiji Adachi

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Contributions

M.E. and Y.S. designed the project and wrote the manuscript. M.E., N.T., M.K. and E.S performed experiments. K.S. provided critical technical information on matrix experiments. H.I., S.O. and T.A. performed computer simulation by discussing details with M.E. and Y.S.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Mototsugu Eiraku or Yoshiki Sasai.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-6 with legends and Legends for Supplementary Movies 1-8.

Videos

  1. 1.

    Supplementary Movie 1

    This movie shows evagination of Rx+ vesicles from an SFEBq-cultured ESC aggregate.

  2. 2.

    Supplementary Movie 2

    This movie shows eye-cup morphogenesis of ESC-derived retinal tissues in 3D live imaging

  3. 3.

    Supplementary Movie 3

    This movie shows inhibition of invagination by aphidicolin treatment.

  4. 4.

    Supplementary Movie 4

    This movie shows tissue dynamics responses to 3D-pinpointed cell ablation by multi photon laser.

  5. 5.

    Supplementary Movie 5

    This movie shows invagination in the ESC-derived retinal epithelium isolated and cocultured with Wnt-expressing cells.

  6. 6.

    Supplementary Movie 6

    This movie shows interkinetic nuclear migration in ESC-derived NR tissues.

  7. 7.

    Supplementary Movie 7

    This movie shows eversion of the RPE-hinge portion of the Phase-4 cup occurring after excision at the proximal hinge.

  8. 8.

    Supplementary Movie 8

    This movie shows computer-simulated animation of the invagination process.

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DOI

https://doi.org/10.1038/nature09941

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