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Dental Pulp Stem Cells: Isolation, Characterization, Expansion, and Odontoblast Differentiation for Tissue Engineering

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Odontogenesis

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

Abstract

Tissue engineering is an interdisciplinary area offering a promising approach by the use of stem cells combined with scaffolds and signaling factors for regeneration of damaged or lost tissues. Incorporation of a sufficient number of cells which do not elicit the immunoreaction in the body is a pivotal element for successful tissue formation using this method. Stem cells exhibiting strong capacity to self-renew and differentiate into different cell types are considered as a potent cell source. Among various cell sources, dental pulp stem cells (DPSCs) are widely under investigation due to the fact that they are simply obtainable from extracted third molars or orthodontically extracted teeth and show an excellent potential for clinical application and also their harvesting method is minimally invasive. DPSCs are odontogenic progenitor cells with clonogenic abilities, rapid proliferation rates, and multiple differentiation potentials. Here, we describe protocols that allow 1) the isolation of DPSCs from a single tooth; 2) the characterization of human mesenchymal stem cells markers of DPSCs by flow cytometry; 3) the culture growth of DPSCs in 2D (in cell culture flasks) and 3D (by 3D printing of cell-laden constructs); and 4) the in vivo evaluation of differentiation potential of DPSCs.

Qing Dong and Yuanyuan Wang contributed equally to this work

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References

  1. Lanza R, Langer R, Vacanti JP (2011) Principles of tissue engineering. Academic Press, Cambridge, MA

    Google Scholar 

  2. Lee SJ, Yoo JJ, Atala A (2018) Biomaterials and tissue engineering. In: Clinical regenerative medicine in urology. Springer, Berlin, pp 17–51

    Chapter  Google Scholar 

  3. Park J et al (2017) Fabrication and characterization of 3D-printed bone-like β-tricalcium phosphate/polycaprolactone scaffolds for dental tissue engineering. J Ind Eng Chem 46:175–181

    Article  CAS  Google Scholar 

  4. Ning L, Chen X (2017) A brief review of extrusion-based tissue scaffold bio-printing. Biotechnol J 12

    Google Scholar 

  5. Lutolf MP, Hubbell JA (2005) Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol 23(1):47

    Article  CAS  Google Scholar 

  6. Assanah F, Khan Y (2018) Cell responses to physical forces, and how they inform the design of tissue-engineered constructs for bone repair: a review. J Mater Sci 53(8):5618–5640

    Article  CAS  Google Scholar 

  7. Heath CA (2000) Cells for tissue engineering. Trends Biotechnol 18(1):17–19

    Article  CAS  Google Scholar 

  8. Bianco P, Robey PG (2001) Stem cells in tissue engineering. Nature 414(6859):118

    Article  CAS  Google Scholar 

  9. Atala A, Lanza R, Thomson JA, Nerem R (2010) Principles of regenerative medicine. Academic Press, Cambridge, MA

    Google Scholar 

  10. Chalisserry EP, Nam SY, Park SH, Anil S (2017) Therapeutic potential of dental stem cells. J Tissue Eng 8:2041731417702531

    Article  Google Scholar 

  11. 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 97(25):13625–13630

    Article  CAS  Google Scholar 

  12. Sharpe PT (2016) Dental mesenchymal stem cells. Development 143(13):2273–2280

    Article  CAS  Google Scholar 

  13. Demirci S, Doğan A, Şahin F (2016) Dental stem cells vs. other mesenchymal stem cells: their pluripotency and role in regenerative medicine. In: Dental stem cells. Springer, Berlin, pp 109–124

    Chapter  Google Scholar 

  14. Nuti N, Corallo C, Chan BMF, Ferrari M, Gerami-Naini B (2016) Multipotent differentiation of human dental pulp stem cells: a literature review. Stem Cell Rev Rep 12(5):511–523

    Article  CAS  Google Scholar 

  15. Tatullo M, Marrelli M, Shakesheff KM, White LJ (2015) Dental pulp stem cells: function, isolation and applications in regenerative medicine. J Tissue Eng Regen Med 9(11):1205–1216

    Article  Google Scholar 

  16. Sunil PM, Manikandan R, Muthumurugan TRY, Sivakumar M (2015) Harvesting dental stem cells-overview. J Pharm Bioallied Sci 7(Suppl 2):S384

    Article  CAS  Google Scholar 

  17. Bakkar M et al (2017) A simplified and systematic method to isolate, culture, and characterize multiple types of human dental stem cells from a single tooth. Methods Mol Biol 1553:191–207

    Article  CAS  Google Scholar 

  18. Cvikl B et al (2017) Response of human dental pulp cells to a silver-containing PLGA/TCP-nanofabric as a potential antibacterial regenerative pulp-capping material. BMC Oral Health 17(1):57

    Article  Google Scholar 

  19. Sakai VT, Cordeiro MM, Dong Z, Zhang Z, Zeitlin BD, Nör JE (2011) Tooth slice/scaffold model of dental pulp tissue engineering. Adv Dent Res 23(3):325–332

    Article  CAS  Google Scholar 

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Correspondence to Petros Papagerakis .

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Dong, Q. et al. (2019). Dental Pulp Stem Cells: Isolation, Characterization, Expansion, and Odontoblast Differentiation 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_9

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  • DOI: https://doi.org/10.1007/978-1-4939-9012-2_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9011-5

  • Online ISBN: 978-1-4939-9012-2

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