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Generation of a Bioengineered Lacrimal Gland by Using the Organ Germ Method

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1597)

Abstract

In organogenesis including lacrimal gland development, cell arrangement within a tissue plays an important role. The lacrimal gland develops from embryonic ocular surface epithelium through reciprocal epithelial and mesenchymal interaction, which is organized by interactive regulation of various pathways of signaling molecules. Current development of an in vitro three-dimensional cell manipulation procedure to generate a bioengineered organ germ, named as the organ germ method, has shown the regeneration of a histologically correct and fully functional bioengineered lacrimal gland after engraftment in vivo. This method demonstrated a possibility of lacrimal gland organ replacement to treat dry eye disease, which has been a public health problem leading reduction of visual function. Here, we describe protocols for lacrimal gland germ regeneration using the organ germ method and methods for analyzing the function of the bioengineered lacrimal gland after its transplantation in vivo.

Key words

Bioengineered lacrimal gland Organ germ method Cell manipulation Transplantation Lacrimal gland germ 

References

  1. 1.
    Schechter JE, Warren DW, Mircheff AK (2010) A lacrimal gland is a lacrimal gland, but rodent’s and rabbit’s are not human. Ocul Surf 8(3):111–134CrossRefPubMedGoogle Scholar
  2. 2.
    Makarenkova HP et al (2000) FGF10 is an inducer and Pax6 a competence factor for lacrimal gland development. Development 127(12):2563–2572PubMedGoogle Scholar
  3. 3.
    Mishima S (1965) Some physiological aspects of the precorneal tear film. Arch Ophthalmol 73:233–241CrossRefPubMedGoogle Scholar
  4. 4.
    Dean C, Ito M, Makarenkova HP, Faber SC, Lang RA (2004) Bmp7 regulates branching morphogenesis of the lacrimal gland by promoting mesenchymal proliferation and condensation. Development 131(17):4155–4165CrossRefPubMedGoogle Scholar
  5. 5.
    Tsau C et al (2011) Barx2 and Fgf10 regulate ocular glands branching morphogenesis by controlling extracellular matrix remodeling. Development 138(15):3307–3317CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Brockes JP, Kumar A (2005) Appendage regeneration in adult vertebrates and implications for regenerative medicine. Science 310(5756):1919–1923CrossRefPubMedGoogle Scholar
  7. 7.
    Atala A (2005) Tissue engineering, stem cells and cloning: current concepts and changing trends. Expert Opin Biol Ther 5(7):879–892CrossRefPubMedGoogle Scholar
  8. 8.
    Qu X et al (2011) Lacrimal gland development and Fgf10-Fgfr2b signaling are controlled by 2-O- and 6-O-sulfated heparan sulfate. J Biol Chem 286(16):14435–14444CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Zoukhri D (2010) Mechanisms involved in injury and repair of the murine lacrimal gland: role of programmed cell death and mesenchymal stem cells. Ocul Surf 8(2):60–69CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Lin H et al (2015) Three-dimensional culture of functional adult rabbit lacrimal gland epithelial cells on decellularized scaffold. Tissue Eng Part A 22(1–2):65–74PubMedGoogle Scholar
  11. 11.
    Nakao K et al (2007) The development of a bioengineered organ germ method. Nat Methods 4(3):227–230CrossRefPubMedGoogle Scholar
  12. 12.
    Ikeda E et al (2009) Fully functional bioengineered tooth replacement as an organ replacement therapy. Proc Natl Acad Sci U S A 106(32):13475–13480CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Toyoshima KE et al (2012) Fully functional hair follicle regeneration through the rearrangement of stem cells and their niches. Nat Commun 3:784CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Hirayama M et al (2013) Functional lacrimal gland regeneration by transplantation of a bioengineered organ germ. Nat Commun 4:2497PubMedPubMedCentralGoogle Scholar
  15. 15.
    Ogawa M et al (2013) Functional salivary gland regeneration by transplantation of a bioengineered organ germ. Nat Commun 4:2498CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Hirayama M, Oshima M, Tsuji T (2013) Development and prospects of organ replacement regenerative therapy. Cornea 32(Suppl 1):S13–S21CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of OphthalmologyKeio University School of MedicineShinjuku-kuJapan
  2. 2.Department of OphthalmologyKeio University School of MedicineShinjuku-kuJapan
  3. 3.Laboratory for Organ RegenerationRIKEN Center for Developmental BiologyKobeJapan
  4. 4.Organ Technologies Inc., Chiyoda-kuTokyoJapan

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