Hair Follicle Regeneration by Transplantation of a Bioengineered Hair Follicle Germ

  • Katsunari Tezuka
  • Koh-ei Toyoshima
  • Takashi TsujiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1453)


Hair follicle morphogenesis is first induced by epithelial–mesenchymal interactions in the developing embryo. In the hair follicle, various stem-cell populations are maintained in specialized niches to promote repetitive hair follicle-morphogenesis, which is observed in the variable lower region of the hair follicle as a postnatal hair cycle. In contrast, the genesis of most organs is induced only once during embryogenesis. We developed a novel bioengineering technique, the Organ Germ Method, that employs three-dimensional stem cell culture for regenerating various organs and reproducing embryonic organogenesis. In this chapter, we describe a protocol for hair follicle germ reconstitution using adult follicle-derived epithelial stem cells and dermal papilla cells with intracutaneous transplantation of the bioengineered hair-follicle organ germ. This protocol can be useful not only for the clinical study of hair regeneration but also for studies of stem cell biology and organogenesis.

Key words

Hair follicle Organ Germ Method Stem cells Epithelial–mesenchymal interaction 


  1. 1.
    Schneider MR et al (2009) The hair follicle as a dynamic miniorgan. Curr Biol 19:132–142CrossRefGoogle Scholar
  2. 2.
    Hardy MH (1992) The secret life of the hair follicle. Trends Genet 8:55–61CrossRefPubMedGoogle Scholar
  3. 3.
    Oshima H et al (2001) Morphogenesis and renewal of hair follicles from adult multipotent stem cells. Cell 104:233–245CrossRefPubMedGoogle Scholar
  4. 4.
    Blanpain C et al (2004) Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. Cell 118:635–648CrossRefPubMedGoogle Scholar
  5. 5.
    Jahoda CA et al (2003) Hair follicle dermal cells differentiate into adipogenic and osteogenic lineages. Exp Dermatol 12:849–859CrossRefPubMedGoogle Scholar
  6. 6.
    Stenn K et al (2007) Bioengineering the hair follicle. Organogenesis 3:6–13CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Nishimura EK et al (2002) Dominant role of the niche in melanocyte stem-cell fate determination. Nature 416:854–860CrossRefPubMedGoogle Scholar
  8. 8.
    Nishimura EK et al (2005) The incomplete melanocyte stem cell maintenance in the niche. Science 4:720–724CrossRefGoogle Scholar
  9. 9.
    Fujiwara H et al (2011) The basement membrane of hair follicle stem cells is a muscle cell niche. Cell 144:577–589CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Sharpe PT, Young CS (2005) Test-tube teeth. Sci Am 293:34–41CrossRefPubMedGoogle Scholar
  11. 11.
    Sasai Y (2013) Cytosystems dynamics in self-organization of tissue architecture. Nature 493:318–326CrossRefPubMedGoogle Scholar
  12. 12.
    Jahoda CA et al (1984) Induction of hair growth by implantation of cultured dermal papilla cells. Nature 311:560–562CrossRefPubMedGoogle Scholar
  13. 13.
    McElwee KJ et al (2003) Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol 121:1267–1275CrossRefPubMedGoogle Scholar
  14. 14.
    Weinberg WC et al (1993) Reconstitution of hair follicle development in vivo: determination of follicle formation, hair growth, and hair quality by dermal cells. J Invest Dermatol 100:229–236CrossRefPubMedGoogle Scholar
  15. 15.
    Takeda A et al (1998) Histodifferentiation of hair follicles in grafting of cell aggregates obtained by rotation culture of embryonic rat skin. Scand J Plast Reconstr Surg Hand Surg 32:359–364CrossRefPubMedGoogle Scholar
  16. 16.
    Lichti U et al (2008) Isolation and short-term culture of primary keratinocytes, hair follicle populations and dermal cells from newborn mice and keratinocytes from adult mice for in vitro analysis and for grafting to immunodeficient mice. Nat Protoc 3:799–810CrossRefPubMedGoogle Scholar
  17. 17.
    Chuong CM et al (2007) Defining hair follicles in the age of stem cell bioengineering. J Invest Dermatol 127:2098–2100CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Nakao K et al (2007) The development of a bioengineered organ germ method. Nat Methods 4:227–230CrossRefPubMedGoogle Scholar
  19. 19.
    Ikeda E et al (2009) Fully functional bioengineered hair replacement as an organ replacement therapy. Proc Natl Acad Sci U S A 106:13475–13480CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Ogawa M et al (2013) Fully functional salivary gland regeneration by transplantation of a bioengineered organ germ. Nat Commun. 784. doi: 10.1038/ncomms3498
  21. 21.
    Hirayama M et al (2013) Fully functional lacrimal gland regeneration by transplantation of a bioengineered organ germ. Nat Commun. doi: 10.1038/ncomms3498
  22. 22.
    Toyoshima K et al (2012) Fully functional hair follicle regeneration through the rearrangement of stem cells and their niches. Nat Commun 3:784CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Asakawa K et al (2012) Hair organ regeneration via the bioengineered hair follicular unit transplantation. Sci Rep 2:424CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Iida M et al (2007) Hair cycle-dependent changes of alkaline phosphatase activity in the mesenchyme and epithelium in mouse vibrissal follicles. Dev Growth Differ 49:185–195CrossRefPubMedGoogle Scholar
  25. 25.
    Osada A et al (2007) Long-term culture of mouse vibrissal dermal papilla cells and de novo hair follicle induction. Tissue Eng 13:975–982CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Katsunari Tezuka
    • 1
    • 2
  • Koh-ei Toyoshima
    • 1
    • 2
    • 3
  • Takashi Tsuji
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Center for Developmental BiologyRIKENKobeJapan
  2. 2.Organ Technologies Inc.TokyoJapan
  3. 3.Department of Regenerative Medicine, Plastic and Reconstructive SurgeryKitasato University of MedicineSagamiharaJapan
  4. 4.Research Institute for Science and TechnologyTokyo University of ScienceNodaJapan

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