Isolation of Dental Stem Cell-Enriched Populations from Continuously Growing Mouse Incisors

  • Anamaria Balic
Part of the Methods in Molecular Biology book series (MIMB, volume 1922)


Continuous growth of the rodent incisor is enabled by epithelial and mesenchymal stem cells (ESCs and MSCs) which unceasingly replenish enamel and dentin, respectively, that wear by persistent animal gnawing. Lineage tracing studies have provided evidence that ESCs contribute to all epithelial lineages of the tooth in vivo. Meanwhile, in the mouse incisor, MSCs continuously contribute to odontoblast lineage and tooth growth. However, in vitro manipulation of ESCs has shown little progress, mainly due to lack of appropriate protocol to successfully isolate, culture, expand, and differentiate ESCs in vitro without using the co-culture system. In this chapter we describe the isolation of the Sox2-GFP+ cell population that is highly enriched in ESCs. Isolated cells can be used for various types of analyses, including in vitro culture, single cell-related analyses, etc. Furthermore, we describe ways to obtain populations enriched in the incisor MSCs using FACS sorting of antibody-labeled cells. Easily accessible FACS sorting enables easy and relatively fast isolation of the cells labeled by the fluorescent protein.

Key words

Dental tissues Stem cell-enriched population Mouse incisors Tooth regeneration 


  1. 1.
    Balic A, Thesleff I (2015) Tissue interactions regulating tooth development and renewal. Curr Top Dev Biol 115:157–186CrossRefGoogle Scholar
  2. 2.
    Harada H, Kettunen P, Jung HS, Mustonen T, Wang YA, Thesleff I (1999) Localization of putative stem cells in dental epithelium and their association with notch and fgf signaling. J Cell Biol 147(1):105–120CrossRefGoogle Scholar
  3. 3.
    Li L, Kwon HJ, Harada H, Ohshima H, Cho SW, Jung HS (2011) Expression patterns of abcg2, bmi-1, oct-3/4, and yap in the developing mouse incisor. Gene Expr Patterns: GEP 11(3–4):163–170CrossRefGoogle Scholar
  4. 4.
    Suomalainen M, Thesleff I (2010) Patterns of wnt pathway activity in the mouse incisor indicate absence of wnt/beta-catenin signaling in the epithelial stem cells. Dev Dyn 239(1):364–372PubMedGoogle Scholar
  5. 5.
    Biehs B, Hu JK, Strauli NB, Sangiorgi E, Jung H, Heber RP, Ho S, Goodwin AF, Dasen JS, Capecchi MR et al (2013) Bmi1 represses ink4a/arf and hox genes to regulate stem cells in the rodent incisor. Nat Cell Biol 15(7):846–852CrossRefGoogle Scholar
  6. 6.
    Juuri E, Saito K, Ahtiainen L, Seidel K, Tummers M, Hochedlinger K, Klein OD, Thesleff I, Michon F (2012) Sox2+ stem cells contribute to all epithelial lineages of the tooth via sfrp5+ progenitors. Dev Cell 23(2):317–328CrossRefGoogle Scholar
  7. 7.
    Seidel K, Ahn CP, Lyons D, Nee A, Ting K, Brownell I, Cao T, Carano RA, Curran T, Schober M et al (2010) Hedgehog signaling regulates the generation of ameloblast progenitors in the continuously growing mouse incisor. Development 137(22):3753–3761CrossRefGoogle Scholar
  8. 8.
    Balic A, Aguila HL, Caimano MJ, Francone VP, Mina M (2010) Characterization of stem and progenitor cells in the dental pulp of erupted and unerupted murine molars. Bone 46(6):1639–1651CrossRefGoogle Scholar
  9. 9.
    Feng J, Mantesso A, De Bari C, Nishiyama A, Sharpe PT (2011) Dual origin of mesenchymal stem cells contributing to organ growth and repair. Proc Natl Acad Sci U S A 108(16):6503–6508CrossRefGoogle Scholar
  10. 10.
    Kaukua N, Shahidi MK, Konstantinidou C, Dyachuk V, Kaucka M, Furlan A, An Z, Wang L, Hultman I, Ahrlund-Richter L et al (2014) Glial origin of mesenchymal stem cells in a tooth model system. Nature 513(7519):551–554CrossRefGoogle Scholar
  11. 11.
    Zhao H, Feng J, Seidel K, Shi S, Klein O, Sharpe P, Chai Y (2014) Secretion of shh by a neurovascular bundle niche supports mesenchymal stem cell homeostasis in the adult mouse incisor. Cell Stem Cell 14(2):160–173CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Anamaria Balic
    • 1
  1. 1.Research Program in Developmental Biology, Institute of BiotechnologyUniversity of HelsinkiHelsinkiFinland

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