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Self-formation of functional adenohypophysis in three-dimensional culture

Nature volume 480, pages 5762 (01 December 2011) | Download Citation

Abstract

The adenohypophysis (anterior pituitary) is a major centre for systemic hormones. At present, no efficient stem-cell culture for its generation is available, partly because of insufficient knowledge about how the pituitary primordium (Rathke’s pouch) is induced in the embryonic head ectoderm. Here we report efficient self-formation of three-dimensional adenohypophysis tissues in an aggregate culture of mouse embryonic stem (ES) cells. ES cells were stimulated to differentiate into non-neural head ectoderm and hypothalamic neuroectoderm in adjacent layers within the aggregate, and treated with hedgehog signalling. Self-organization of Rathke’s-pouch-like three-dimensional structures occurred at the interface of these two epithelia, as seen in vivo, and various endocrine cells including corticotrophs and somatotrophs were subsequently produced. The corticotrophs efficiently secreted adrenocorticotropic hormone in response to corticotrophin releasing hormone and, when grafted in vivo, these cells rescued the systemic glucocorticoid level in hypopituitary mice. Thus, functional anterior pituitary tissue self-forms in ES cell culture, recapitulating local tissue interactions.

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Acknowledgements

We are grateful to H. Enomoto, R. Ladher and M. Eiraku for invaluable comments, to K. Misaki for electron microscopy analysis, and to members of the Y.S. laboratory for discussion. This work was supported by grants-in-aid from Ministry of Education, Culture, Sports, Science and Technology (Y.S., Y.O.), the Knowledge Cluster Initiative at Kobe, and the Leading Project for Realization of Regenerative Medicine (Y.S.).

Author information

Affiliations

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

    • Hidetaka Suga
    • , Taisuke Kadoshima
    • , Maki Minaguchi
    • , Mika Soen
    • , Tokushige Nakano
    • , Nozomu Takata
    • , Takafumi Wataya
    • , Keiko Muguruma
    •  & Yoshiki Sasai
  2. Division of Human Stem Cell Technology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Hidetaka Suga
    •  & Masatoshi Ohgushi
  3. Department of Endocrinology and Diabetes, Graduate School of Medicine, Nagoya University, Nagoya, 466-8550, Japan

    • Hidetaka Suga
    •  & Yutaka Oiso
  4. Subteam for Manipulation of Cell Fate, BioResource Center, RIKEN, Tsukuba 305-0074, Japan

    • Hiroyuki Miyoshi
  5. Electron Microscopy Laboratory, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Shigenobu Yonemura

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Contributions

H.S. and Y.S. designed the project and wrote the manuscript. H.S., T.K., M.O. and M.M. performed the experiments with the technical help and advice of T.N., N.T., M.S., K.M., H.M., S.Y. and T.W., and Y.O. provided critical advice on the research strategy and design.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Yoshiki Sasai.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-9 with legends and legends for Supplementary Movies 1-2.

Videos

  1. 1.

    Supplementary Movie 1

    This movie shows the formation of Lim3+ vesicles in SFEBq-cultured ESC Aggregates - see Supplementary Information file for full legend.

  2. 2.

    Supplementary Movie 2

    This movie shows improved locomotor activity in hypophysectomized mice receiving SAG+DAPT-treated aggregates - see Supplementary Information file for full legend.

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DOI

https://doi.org/10.1038/nature10637

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