Skip to main content
SpringerLink
Log in

Dental Mesenchymal Stem Cells: Dental Pulp Stem Cells, Periodontal Ligament Stem Cells, Apical Papilla Stem Cells, and Primary Teeth Stem Cells—Isolation, Characterization, and Expansion for Tissue Engineering

  • Protocol
  • First Online:
Odontogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1922))

Abstract

Dental stem cells (DSCs) have been shown to possess great potential for multiple biomedical applications, especially for dental tissue regeneration. They are a special type of subpopulation of mesenchymal stem/stromal cells (MSCs) and present subtle differences from other types of MSCs. Therefore, it requires a specialized expertise to isolate, culture, and characterize these cells in vitro and in vivo. The purpose of this chapter is to share our experience in studying these cells. We will describe in detail laboratory protocols outlining how the cells are isolated, cultured, expanded, and characterized using various in vitro cellular and biochemical analyses, as well as an in vivo study model using immunocompromised mice to observe tissue regeneration after transplantation of these DSCs.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Huang GTJ, Gronthos S, Shi S (2009) Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res 88(9):792–806

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Morsczeck C, Huang GT-J, Shi S (2013) Stem and progenitor cells of dental and gingival tissue origin. In: Huang GT-J, Thesleff I (eds) Stem cells, craniofacial development and regeneration, 1st edn. Wiley-Blackwell, Hoboken, NJ, pp 285–302

    Google Scholar 

  3. Huang GTJ, Garcia-Godoy F (2017) Stem cells and dental tissue reconstruction. In: Spencer P, Misra A (eds) Material-tissue interfacial phenomena, 1st edn. Elsevier, Woodhead Publishing, Sawston, Cambridge

    Google Scholar 

  4. Huang GTJ, Yamaza T, Shea LD, Djouad F, Kuhn NZ, Tuan RS et al (2009) Stem/progenitor cell-mediated de novo regeneration of dental pulp with newly deposited continuous layer of dentin in an in vivo model. Tissue Eng Part A 16(2):605–615

    PubMed Central  Google Scholar 

  5. Nakashima M, Huang GT-J (2013) Pulp and dentin regeneration. In: Huang GT-J, Thesleff I (eds) Stem cells in craniofacial development and regeneration, 1st edn. Wiley-Blackwell, Hoboken, NJ, pp 461–484

    Google Scholar 

  6. Ishizaka R, Hayashi Y, Iohara K, Sugiyama M, Murakami M, Yamamoto T et al (2013) Stimulation of angiogenesis, neurogenesis and regeneration by side population cells from dental pulp. Biomaterials 34(8):1888–1897

    CAS  PubMed  Google Scholar 

  7. Iohara K, Zheng L, Wake H, Ito M, Nabekura J, Wakita H et al (2008) A novel stem cell source for vasculogenesis in ischemia: subfraction of side population cells from dental pulp. Stem Cells 26(9):2408–2418

    PubMed  Google Scholar 

  8. Sakai K, Yamamoto A, Matsubara K, Nakamura S, Naruse M, Yamagata M et al (2012) Human dental pulp-derived stem cells promote locomotor recovery after complete transection of the rat spinal cord by multiple neuro-regenerative mechanisms. J Clin Invest 122(1):80–90

    CAS  PubMed  Google Scholar 

  9. Gay IC, Chen S, MacDougall M (2007) Isolation and characterization of multipotent human periodontal ligament stem cells. Orthod Craniofac Res 10(3):149–160

    CAS  PubMed  Google Scholar 

  10. Choi S, Cho TJ, Kwon SK, Lee G, Cho J (2013) Chondrogenesis of periodontal ligament stem cells by transforming growth factor-beta3 and bone morphogenetic protein-6 in a normal healthy impacted third molar. Int J Oral Sci 5(1):7–13

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Yan X, Qin H, Qu C, Tuan RS, Shi S, Huang GT (2010) iPS cells reprogrammed from human mesenchymal-like stem/progenitor cells of dental tissue origin. Stem Cells Dev 19(4):469–480

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Huang GT-J (2010) Induced pluripotent stem cells—a new foundation in medicine. J Exp Clin Med 2(5):202–217

    PubMed  PubMed Central  Google Scholar 

  13. Huang GTJ, El Ayachi I, Zou X-Y (2017) Induced pluripotent stem cell technologies for tissue engineering. In: Waddington RJ, Sloan AJ (eds) Tissue engineering and regeneration in dentistry- current strategies. John Wiley & Sons, Ltd, West Sussex, UK

    Google Scholar 

  14. Gronthos S, Mankani M, Brahim J, Robey PG, Shi S (2000) Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A 97(25):13625–13630

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG et al (2003) SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci U S A 100(10):5807–5812

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J et al (2004) Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 364(9429):149–155

    CAS  PubMed  Google Scholar 

  17. Sonoyama W, Liu Y, Yamaza T, Tuan RS, Wang S, Shi S et al (2008) Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod 34(2):166–171

    PubMed  PubMed Central  Google Scholar 

  18. Morsczeck C, Vollner F, Saugspier M, Brandl C, Reichert TE, Driemel O et al (2010) Comparison of human dental follicle cells (DFCs) and stem cells from human exfoliated deciduous teeth (SHED) after neural differentiation in vitro. Clin Oral Investig 14(4):433–440

    PubMed  Google Scholar 

  19. Zhu X, Liu J, Yu Z, Chen CA, Aksel H, Azim AA, Huang GTJ (2018) A miniature swine model for stem cell-based de novo regeneration of dental pulp and dentin-like tissue. Tissue Eng Part C Methods 24(2):108–120

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Sonoyama W, Liu Y, Fang D, Yamaza T, Seo BM, Zhang C et al (2006) Mesenchymal stem cell-mediated functional tooth regeneration in swine. PLoS One 1:e79

    PubMed  PubMed Central  Google Scholar 

  21. Smith JR, Pochampally R, Perry A, Hsu SC, Prockop DJ (2004) Isolation of a highly clonogenic and multipotential subfraction of adult stem cells from bone marrow stroma. Stem Cells 22(5):823–831

    PubMed  Google Scholar 

  22. Horwitz EM, Le Blanc K, Dominici M, Mueller I, Slaper-Cortenbach I, Marini FC et al (2005) Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. Cytotherapy 7(5):393–395

    CAS  PubMed  Google Scholar 

  23. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini FC, Krause DS et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8(4):315–317

    CAS  PubMed  Google Scholar 

  24. Ebrahimi B, Yaghoobi MM, Kamal-abadi AM, Raoof M (2011) Human dental pulp stem cells express many pluripotency regulators and differentiate into neuronal cells. Neural Regen Res 6(34):2666–2672

    CAS  Google Scholar 

  25. Alongi DJ, Yamaza T, Song Y, Fouad AF, Romberg EE, Shi S et al (2010) Stem/progenitor cells from inflamed human dental pulp retain tissue regeneration potential. Regen Med 5(4):617–631

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Gauthier P, Yu Z, Tran QT, Bhatti FU, Zhu X, Huang GT (2017) Cementogenic genes in human periodontal ligament stem cells are downregulated in response to osteogenic stimulation while upregulated by vitamin C treatment. Cell Tissue Res 368(1):79–92

    CAS  PubMed  Google Scholar 

  27. Yu Z, Gauthier P, Tran QT, El Ayachi I, Bhatti F-U-R, Bahabri R et al (2015) Differential properties of human ALP+ periodontal ligament stem cells vs their ALP- counterparts. J Stem Cell Res Ther 5(7):292

    PubMed  PubMed Central  Google Scholar 

  28. Huang GT, Sonoyama W, Liu Y, Liu H, Wang S, Shi S (2008) The hidden treasure in apical papilla: the potential role in pulp/dentin regeneration and bioroot engineering. J Endod 34(6):645–651

    PubMed  PubMed Central  Google Scholar 

  29. Al-Habib M, Yu Z, Huang GT (2013) Small molecules affect human dental pulp stem cell properties via multiple signaling pathways. Stem Cells Dev 22(17):2402–2413

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported in part by a grant from the US National Institutes of Health R01 DE019156 (G.T.-J.H.), by an Endodontic Research Grant from American Association of Endodontists Foundation (G.T.-J.H.), and by a Research Fund from the University of Tennessee Health Science Center. The authors wish to thank Drs. Philippe Guathier, Zongdong Yu, and Yaa Owusu for their work on PDLSCs.

Author information

Authors and Affiliations

  1. Faculty of Dentistry, Department of Endodontics, King Abdulaziz University, Jeddah, Saudi Arabia

    Mey Al-Habib

  2. Department of Bioscience Research, University of Tennessee Health Science Center, College of Dentistry, Memphis, TN, USA

    George T. -J. Huang

Authors
  1. Mey Al-Habib
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. George T. -J. Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to George T. -J. Huang .

Editor information

Editors and Affiliations

  1. School of Dentistry, University of Michigan, Ann Arbor, MI, USA

    Petros Papagerakis

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Al-Habib, M., Huang, G.T.J. (2019). Dental Mesenchymal Stem Cells: Dental Pulp Stem Cells, Periodontal Ligament Stem Cells, Apical Papilla Stem Cells, and Primary Teeth Stem Cells—Isolation, Characterization, and Expansion for Tissue Engineering. In: Papagerakis, P. (eds) Odontogenesis. Methods in Molecular Biology, vol 1922. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9012-2_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9012-2_7

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9011-5

  • Online ISBN: 978-1-4939-9012-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation