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Melanoblasts as Multipotent Cells in Murine Skin

  • Tsutomu MotohashiEmail author
  • Takahiro Kunisada
Part of the Methods in Molecular Biology book series


Melanoblasts (MBs) are melanocyte precursors that are derived from neural crest cells (NCCs). We recently demonstrated the multipotency of MBs; they differentiate not only into pigmented melanocytes but also other NCC derivatives. We herein describe methods for the isolation of MBs from mouse skin by flow cytometry. Methods to culture isolated MBs that retain their multipotency and isolation methods for MBs using gene-modified mouse are also described.


CD45 Flow cytometer KIT Melanoblasts Multipotency SOX10 



This study was supported by a grant from the program Grants-in-Aid for Scientific Research (C) from the Japan Society for Promotion of Science.


  1. 1.
    Luciani F, Champeval D, Herbette A et al (2011) Biological and mathematical modeling of melanocyte development. Development 138:3943–3954Google Scholar
  2. 2.
    Larue L, de Vuyst F, Delmas V (2013) Modeling melanoblast development. Cell Mol Life Sci 70:1067–1079Google Scholar
  3. 3.
    Le Douarin NM, Kalcheim C (1999) The neural crest, 2nd edn. Cambridge University Press, CambridgeGoogle Scholar
  4. 4.
    Wilson YM, Richards KL, Ford-Perriss ML et al (2004) Neural crest cell lineage segregation in the mouse neural tube. Development 131:6153–6162Google Scholar
  5. 5.
    Fernandes KJ, McKenzie IA, Mill P et al (2004) A dermal niche for multipotent adult skin-derived precursor cells. Nat Cell Biol 6:1082–1093Google Scholar
  6. 6.
    Toma JG, McKenzie IA, Bagli D et al (2005) Isolation and characterization of multipotent skin-derived precursors from human skin. Stem Cells 23:727–737Google Scholar
  7. 7.
    Wong CE, Paratore C, Dours-Zimmermann MT et al (2006) Neural crest-derived cells with stem cell features can be traced back to multiple lineages in the adult skin. J Cell Biol 175:1005–1015Google Scholar
  8. 8.
    Sieber-Blum M, Grim M, Hu YF et al (2004) Pluripotent neural crest stem cells in the adult hair follicle. Dev Dyn 231:258–269Google Scholar
  9. 9.
    Amoh Y, Li L, Katsuoka K et al (2005) Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. Proc Natl Acad Sci U S A 102:5530–5534Google Scholar
  10. 10.
    Yu H, Fang D, Kumar SM et al (2006) Isolation of a novel population of multipotent adult stem cells from human hair follicles. Am J Pathol 168:1879–1888Google Scholar
  11. 11.
    Dupin E, Glavieux C, Vaigot P et al (2000) Endothelin 3 induces the reversion of melanocytes to glia through a neural crest-derived glial-melanocytic progenitor. Proc Natl Acad Sci U S A 97:7882–7887Google Scholar
  12. 12.
    Trentin A, Glavieux-Pardanaud C, Le Douarin NM et al (2004) Self-renewal capacity is a widespread property of various types of neural crest precursor cells. Proc Natl Acad Sci U S A 101:4495–4500Google Scholar
  13. 13.
    Real C, Glavieux-Pardanaud C, Le Douarin NM et al (2006) Clonally cultured differentiated pigment cells can dedifferentiate and generate multipotent progenitors with self-renewing potential. Dev Biol 300:656–669Google Scholar
  14. 14.
    Dupin E, Real C, Glavieux-Pardanaud C et al (2003) Reversal of developmental restrictions in neural crest lineages: transition from Schwann cells to glial-melanocytic precursors in vitro. Proc Natl Acad Sci U S A 100:5229–5233Google Scholar
  15. 15.
    Real C, Glavieux-Pardanaud C, Vaigot P et al (2005) The instability of the neural crest phenotypes: Schwann cells can differentiate into myofibroblasts. Int J Dev Biol 49:151–159Google Scholar
  16. 16.
    Motohashi T, Yamanaka K, Chiba K et al (2009) Unexpected multipotency of melanoblasts isolated from murine skin. Stem Cells 27:888–897Google Scholar
  17. 17.
    Lagasse E, Connors H, Al-Dhalimy M et al (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med 6:1229–1234Google Scholar
  18. 18.
    Motohashi T, Yamanaka K, Chiba K et al (2011) Neural crest cells retain their capability for multipotential differentiation even after lineage-restricted stages. Dev Dyn 240:1681–1693Google Scholar
  19. 19.
    Nagai T, Ibata K, Park ES et al (2002) A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol 20:87–90Google Scholar
  20. 20.
    Motohashi T, Kunisada T (2018) Direct conversion of mouse embryonic fibroblasts into neural crest cells. Methods Mol Biol.
  21. 21.
    Rugh R (1990) The mouse. Oxford University Press, New YorkGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Department of Tissue and Organ Development, Regeneration and Advanced Medical ScienceGifu University Graduate School of MedicineGifuJapan

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