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Isolation and in vitro characterisation of dental pulp stem cells from natal teeth

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Abstract

Dental pulp stem cells were primarily derived from the pulp tissues of exfoliated deciduous teeth, primary incisors and permanent third molar teeth. The aim of this study was to isolate and extensively characterise SCs derived from human natal dental pulp (hNDP). For characterisation, proliferation capacity, phenotypic properties, ultrastructural and differentiation characteristics and gene expression profiles were utilised. A comparison was done between the properties of NDP-SCs and the properties of mesenchymal stem cells (MSCs) from bone marrow (BM) of the human. Stem cells isolated from hNDP and hBM were analysed by flow cytometry, reverse transcriptase-PCR, Real Time-PCR, and immunocytochemistry. Both cell lines were directionally differentiated towards adipogenic, osteogenic chondrogenic, myogenic and neurogenic lineages. hNDP-SCs and hBM-MSCs expressed CD13, CD44, CD90, CD146 and CD166, but not CD3, CD8, CD11b, CD14, CD15, CD19, CD33, CD34, CD45, CD117, and HLA-DR. Ultrastructural characteristics of hNDP-SCs showed more developed and metabolically active cells. hNDP-SCs and hBM-MSCs expressed some adipogenic (leptin, adipophilin and PPARγ), myogenic (desmin, myogenin, myosinIIa, and α-SMA), neurogenic (γ-enolase, MAP2a,b, c-fos, nestin, NF-H, NF-L, GFAP and betaIII tubulin), osteogenic (osteonectin, osteocalcin, osteopontin, Runx-2, and type I collagen) and chondrogenic (type II collagen, SOX9) markers without any stimulation towards differentiation under basal conditions. Embryonic stem cell markers Oct4, Rex-1, FoxD-3, Sox2, and Nanog were also identified. The differentiation potential of hNDP-SCs and hBM-MSCs to adipogenic, osteogenic, chondrogenic, myogenic and neurogenic was shown. This report described the first successful isolation and characterisation of hNDP-SCs.

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References

  • Arthur A, Rychkov G, Shi S, Koblar SA, Gronthos S (2008) Adult human dental pulp stem cells differentiate toward functionally active neurons under appropriate environmental cues. Stem Cells 26:1787–1795

    Article  CAS  PubMed  Google Scholar 

  • Bjerre L, Bunger CE, Kassem M, Mygind T (2008) Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds. Biomaterials 29:2616–2627

    Article  CAS  PubMed  Google Scholar 

  • d’Aquino R, Graziano A, Sampaolesi M, Laino G, Pirozzi G, De Rosa A, Papaccio G (2007) Human postnatal dental pulp cells co-differentiate into osteoblasts and endotheliocytes: a pivotal synergy leading to adult bone tissue formation. Cell Death Differ 14:1162–1171

    Article  PubMed  Google Scholar 

  • Gandia C, Armiñan A, García-Verdugo JM et al (2008) Human dental pulp stem cells improve left ventricular function, induce angiogenesis, and reduce infarct size in rats with acute myocardial infarction. Stem Cells 26:638–645

    Article  PubMed  Google Scholar 

  • 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 USA 97:13625–13630

    Article  CAS  PubMed  Google Scholar 

  • Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, DenBesten P, Robey PG, Shi S (2002) Stem cell properties of human dental pulp stem cells. J Dent Res 81:531–535

    Article  CAS  PubMed  Google Scholar 

  • Huang AH, Chen YK, Lin LM, Shieh TY, Chan AW (2008) Isolation and characterization of dental pulp stem cells from a supernumerary tooth. J Oral Pathol Med 37:571–574

    PubMed  Google Scholar 

  • Ikeda E, Hirose M, Kotobuki N, Shimaoka H, Tadokoro M, Maeda M, Hayashi Y, Kirita T, Ohgushi H (2006) Osteogenic differentiation of human dental papilla mesenchymal cells. Biochem Biophys Res Commun 342:1257–1262

    Article  CAS  PubMed  Google Scholar 

  • Iohara K, Nakashima M, Ito M, Ishikawa M, Nakasima A, Akamine A (2004) Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2. J Dent Res 83:590–595

    Article  CAS  PubMed  Google Scholar 

  • Izadpanah R, Trygg C, Patel B, Kriedt C, Dufour J, Gimble JM, Bunnell BA (2006) Biologic properties of mesenchymal stem cells derived from bone marrow and adipose tissue. J Cell Biochem 99:1285–1297

    Article  CAS  PubMed  Google Scholar 

  • Jo YY, Lee HJ, Kook SY, Choung HW, Park JY, Chung JH, Choung YH, Kim ES, Yang HC, Choung PH (2007) Isolation and characterization of postnatal stem cells from human dental tissues. Tissue Eng 13:767–773

    Article  CAS  PubMed  Google Scholar 

  • Kerkis I, Kerkis A, Dozortsev D, Stukart-Parsons GC, Gomes Massironi SM, Pereira LV, Caplan AI, Cerruti HF (2006) Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cell markers. Cells Tissues Organs 184:105–116

    Article  CAS  PubMed  Google Scholar 

  • Laino G, d’Aquino R, Graziano A, Lanza V, Carinci F, Naro F, Pirozzi G, Papaccio G (2005) A new population of human adult dental pulp stem cells: a useful source of living autologous fibrous bone tissue (LAB). J Bone Miner Res 20:1394–1402

    Article  PubMed  Google Scholar 

  • Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG, Shi S (2003) SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA 100:5807–5812

    Article  CAS  PubMed  Google Scholar 

  • Myginda T, Stiehler M, Baatrup A, Li H, Zou X, Flyvbjerg A, Kassem M, Bünger C (2007) Mesenchymal stem cell ingrowth and differentiation on coralline hydroxyapatite scaffolds. Biomaterials 28:1036–1047

    Article  Google Scholar 

  • Nosrat IV, Widenfalk J, Olson L, Nosrat CA (2001) Dental pulp cells produce neurotrophic factors, interact with trigeminal neurons in vitro, and rescue motoneurons after spinal cord injury. Dev Biol 238:120–132

    Article  CAS  PubMed  Google Scholar 

  • Otaki S, Ueshima S, Shiraishi K, Sugiyama K, Hamada S, Yorimoto M, Matsuo O (2007) Mesenchymal progenitor cells in adult human dental pulp and their ability to form bone when transplanted into immunocompromised mice. Cell Biol Int 31:1191–1197

    Article  CAS  PubMed  Google Scholar 

  • Papaccio G, Graziano A, d’Aquino R, Graziano MF, Pirozzi G, Menditti D, De Rosa A, Carinci F, Laino G (2006) Long-term cryopreservation of dental pulp stem cells (SBP-DPSCs) and their differentiated osteoblasts: a cell source for tissue repair. J Cell Physiol 208:319–325

    Article  CAS  PubMed  Google Scholar 

  • Serakinci N, Graakjaer J, Kolvraa S (2008) Telomere stability and telomerase in mesenchymal stem cells. Biochimie 90:33–40

    Article  CAS  PubMed  Google Scholar 

  • Shi S, Robey PG, Gronthos S (2001) Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. Bone 29:532–539

    Article  CAS  PubMed  Google Scholar 

  • Sonoyama W, Liu Y, Fang D, Yamaza T, Seo BM, Zhang C, Liu H, Gronthos S, Wang CY, Shi S, Wang S (2006) Mesenchymal stem cell-mediated functional tooth regeneration in swine. PLoS ONE. doi:10.1371/journal.pone.0000079

  • Struys T, Krage T, Martens W, Theunissen E, Moreels M, Lambrichts I (2007) Metallothionein: a possible new marker for human dental pulp stem cells. Eur Cells Mater 14:131

    Google Scholar 

  • Wu L, Wu Y, Lin Y, Jing W, Nie X, Qiao J, Liu L, Tang W, Tian W (2007) Osteogenic differentiation of adipose derived stem cells promoted by overexpression of osterix. Mol Cell Biochem 301:83–92

    Article  CAS  PubMed  Google Scholar 

  • Yu J, Wang Y, Deng Z, Tang L, Li Y, Shi J, Jin Y (2007) Odontogenic capability: bone marrow stromal stem cells versus dental pulp stem cells. Biol Cell 99:465–474

    Article  CAS  PubMed  Google Scholar 

  • Zhang W, Walboomers XF, Shi S, Fan M, Jansen JA (2006a) Multilineage differentiation potential of stem cells derived from human dental pulp after cryopreservation. Tissue Eng 12:2813–2823

    Article  CAS  PubMed  Google Scholar 

  • Zhang W, Walboomers XF, van Kuppevelt TH, Daamen WF, Bian Z, Jansen JA (2006b) The performance of human dental pulp stem cells on different three-dimensional scaffold materials. Biomaterials 27:5658–5668

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgment

We thank Özlem Sağlam and Gülay Bayazıt for her excellent technical assistance.

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Authors and Affiliations

  1. Stem Cell and Gene Therapy Research and Applied Center, Kocaeli University, 41380, Kocaeli, Turkey

    Erdal Karaöz, Ayça Aksoy, Gülçin Gacar, Selda Ayhan, Zehra Seda Genç, Gökhan Duruksu, Pınar Çetinalp Demircan & Ayla Eker Sarıboyacı

  2. Department of Pedodontics, Faculty of Dentistry, University of Marmara, 34100, Istanbul, Turkey

    Burcu Nur Doğan & Serap Akyüz

  3. Department of Histology and Embryology, Faculty of Medicine, University of Hacettepe, 06100, Ankara, Turkey

    Sinan Yürüker

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  1. Erdal Karaöz
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  2. Burcu Nur Doğan
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  3. Ayça Aksoy
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  4. Gülçin Gacar
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  9. Gökhan Duruksu
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Correspondence to Erdal Karaöz.

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Karaöz, E., Doğan, B.N., Aksoy, A. et al. Isolation and in vitro characterisation of dental pulp stem cells from natal teeth. Histochem Cell Biol 133, 95–112 (2010). https://doi.org/10.1007/s00418-009-0646-5

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  • DOI: https://doi.org/10.1007/s00418-009-0646-5

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