Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

The development of a bioengineered organ germ method


To bioengineer ectodermal organs such as teeth and whisker follicles, we developed a three-dimensional organ-germ culture method. The bioengineered tooth germ generated a structurally correct tooth, after both in vitro organ culture as well as transplantation under a tooth cavity in vivo, showing penetration of blood vessels and nerve fibers. Our method provides a substantial advance in the development of bioengineered organ replacement strategies and regenerative therapies. Please visit methagora to view and post comments on this article

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it


Prices may be subject to local taxes which are calculated during checkout

Figure 1: Generation of a whole tooth using bioengineered tooth germ derived from dissociated single cells in vivo.
Figure 2: Development of the bioengineered incisor in the tooth cavity of adult mice.
Figure 3: Analysis of periodontal ligaments, endothelial cells of tooth blood vessels and neural filaments in a bioengineered tooth.


  1. Langer, R.S. & Vacanti, J.P. Sci. Am. 280, 86–89 (1999).

    Article  CAS  Google Scholar 

  2. Brockes, J.P. & Kumar, A. Science 310, 1919–1923 (2005).

    Article  CAS  Google Scholar 

  3. Atala, A. Expert Opin. Biol. Ther. 5, 879–892 (2005).

    Article  CAS  Google Scholar 

  4. Pispa, J. & Thesleff, I. Dev. Biol. 262, 195–205 (2003).

    Article  CAS  Google Scholar 

  5. Thesleff, I. J. Cell Sci. 116, 1647–1648 (2003).

    Article  CAS  Google Scholar 

  6. Tucker, A. & Sharpe, P. Nat. Rev. Genet. 5, 499–508 (2004).

    Article  CAS  Google Scholar 

  7. Sharpe, P.T. & Young, C.S. Sci. Am. 293, 34–41 (2005).

    Article  Google Scholar 

  8. Young, C.S. et al. J. Dent. Res. 81, 695–700 (2002).

    Article  CAS  Google Scholar 

  9. Ohazama, A., Modino, S.A., Miletich, I. & Sharpe, P.T. J. Dent. Res. 83, 518–522 (2004).

    Article  CAS  Google Scholar 

  10. Song, Y. et al. Dev. Dyn. 235, 1334–1344 (2006).

    Article  CAS  Google Scholar 

  11. Hu, B. et al. Tissue Eng. 12, 2069–2075 (2006).

    Article  CAS  Google Scholar 

  12. Steinberg, M.S. Dev. Biol. 180, 377–388 (1996).

    Article  CAS  Google Scholar 

  13. Hata, R.I. Cell Biol. Int. 20, 59–65 (1996).

    Article  CAS  Google Scholar 

  14. Salmivirta, K., Gullberg, D., Hirsch, E., Altruda, F. & Ekblom, P. Dev. Dyn. 205, 104–113 (1996).

    Article  CAS  Google Scholar 

Download references


We thank M. Okabe (Osaka University) for providing the C57BL/6-TgN (act-EGFP) OsbC14-Y01-FM131 mice. We are also grateful to K. Itoh and M. Sugai (Kyoto University) for critical reading of this manuscript. We also thank to T. Katakai (Kyoto University) and Y. Nishi (Nagahama Institute of Bioscience and Technology) for their valuable discussions and encouragement. This work was partially supported by an “Academic Frontier” Project for Private Universities to Y.T. and T.T. (2003–2007) and by a Grant-in Aid for Scientific Research in Priority Areas (50339131) to T.T. from MEXT Japan.

Author information

Authors and Affiliations



K.N. was involved in each of the experiments described in this study. R.M. analyzed the explants that were formed by a combination of normal and GFP-transgenic mouse-derived cells and performed the immunohistochemical analysis shown in Figure 3. Y.S. performed and analyzed the transplantation experiments in both the subrenal capsule and the tooth cavity. K.I. and M.S. performed the in situ hybridization analysis. Y.T. performed the subrenal capsule transplantation experiments and histological analysis. M.O. maintained the in vitro organ cultures and performed whole-mount analysis of the chimeric bioengineered tooth germ. K.N. and T.T. prepared the manuscript. K.N., M.S., Y.T. and T.T. discussed the results and also contributed to the preparation of this manuscript. T.T. designed the experiments.

Corresponding author

Correspondence to Takashi Tsuji.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Effects of cell density and cell compartmentalization between epithelial and mesenchymal cells upon the generation of bioengineered teeth. (PDF 756 kb)

Supplementary Fig. 2

Three-dimensional histological analysis of bioengineered teeth under various developmental conditions. (PDF 644 kb)

Supplementary Fig. 3

Expression of the regulatory genes that function during early tooth development in a bioengineered incisor tooth germ. (PDF 408 kb)

Supplementary Fig. 4

Generation of a reconstituted whisker in vivo from a bioengineered follicle. (PDF 434 kb)

Supplementary Fig. 5

Development and transplantation of individual primordia. (PDF 416 kb)

Supplementary Methods (PDF 27 kb)

Supplementary Note (PDF 24 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nakao, K., Morita, R., Saji, Y. et al. The development of a bioengineered organ germ method. Nat Methods 4, 227–230 (2007).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

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