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Various methods for isolation of multipotent human periodontal ligament cells for regenerative medicine

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Abstract

Periodontal ligament (PDL) is a specialized connective tissue that connects cementum and alveolar bone to maintain and support the teeth in situ and preserve tissue homeostasis. Recent studies have revealed the existence of stem cells in human dental tissues including periodontal ligament that play an important role, not only in the maintenance of the periodontium but also in promoting periodontal regeneration. In this study, human periodontal ligament cells (hPDLCs) were isolated by outgrowth and enzymatic dissociation methods. Expression of surface markers on PDLCs as human mesenchymal stem cells (MSCs) was identified by flow cytometry. In addition, proliferation and differentiation capacity of cultured cells to osteoblasts, adipocytes were evaluated. As a result, we successfully cultured cells from the human periodontal ligament tissues. PDLCs express mesenchymal stem cell (MSC) markers such as CD44, CD73, and CD90 and do not express CD34, CD45, and HLA-DR. PDLCs also possess the multipotential to differentiate into various types of cells, such as osteoblast and adipocytes, in vitro. Therefore, these cells have high potential to serve as materials for tissue engineering, especially dental tissue engineering.

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References

  • Baroffio A, Hamann M, Bernheim L, Bochaton-Piallat ML, Gabbiani G, Bader CR (1996) Identification of self-renewing myoblasts in the progeny of single human muscle satellite cells. Differentiation 60:47–57

    Article  CAS  PubMed  Google Scholar 

  • Bartold PM, Shi S, Gronthos S (2006) Stem cells and periodontal regeneration. Periodontal 40:164–172

    Article  Google Scholar 

  • Batouli S, Miura M, Brahim J, Tsutsui TW, Fisher LW, Gronthos S, Robey PG, Shi S (2003) Comparison of stem cells mediated osteogenesis and dentinogenesis. J Dent Res 82:976–981

    Article  CAS  PubMed  Google Scholar 

  • Campagnoli C, Roberts IA, Kumar S, Bennett PR, Bellantuono I, Fisk NM (2001) Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood 98:2396–2402

    Article  CAS  PubMed  Google Scholar 

  • Feng F, Akiyama K, Liu Y, Yamaza T, Wang TM, Chen JH, Wang BB, Huang GT, Wang S, Shi S (2010) Utility of PDL progenitors for in vivo tissue regeneration: a report of 3 cases. Oral Dis 16:8–20

    Article  Google Scholar 

  • Fujii S, Maeda H, Wada N, Tomokiyo A, Saito M, Akamine A (2008) Investigating a clonal human periodontal ligament progenitor/stem cell line in vitro and in vivo. J Cell Physiol 215:743–749

    Article  CAS  PubMed  Google Scholar 

  • Gay I, Chen S, MacDougall M (2007) Isolation and characterization of multipotent human periodontal ligament stem cells. Orthod Craniofacial Res 10:149–160

    Article  CAS  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 

  • 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 97:13625–13630

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gronthos S, Zannettino AC, Hay SJ, Shi S, Graves SE, Kortesidis A, Simmons PJ (2003) Molecular and cellular characterisation of highly purified stromal stem cells derived from human bone marrow. J Cell Sci 116:1827–1835

    Article  CAS  PubMed  Google Scholar 

  • Handa K, Saito M, Tsunoda A, Yamauchi M, Hattori S, Sato S, Toyoda M, Teranaka T, Narayanan AS (2002) Progenitor cells from dental follicle are able to form cementum matrix in vivo. Connect Tissue Res 43:406–408

    Article  PubMed  Google Scholar 

  • Huang GT, 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 88:792–796

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • 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 34:645–651

    Article  PubMed Central  PubMed  Google Scholar 

  • Kamata N, Fujimoto R, Tomonari M, Taki M, Nagayama M, Yasumoto S (2004) Immortalization of human dental papilla, dental pulp, periodontal ligament cells and gingival fibroblasts by telomerase reverse transcriptase. J Oral Pathol Med 33:417–423

    Article  CAS  PubMed  Google Scholar 

  • Lindroos B, Mäenpää K, Ylikomi T, Oja H, Suuronen R, Miettinen S (2008) Characterisation of human dental stem cells and buccal mucosa fibroblasts. Biochem Biophys Res Commun 368:329–335

    Article  CAS  PubMed  Google Scholar 

  • Marie PJ (2008) Transcription factors controlling osteoblastogenesis. Arch Biochem Biophys 473:98–105

    Article  CAS  PubMed  Google Scholar 

  • McCulloch CA, Nemeth E, Lowenberg B, Melcher AH (1987) Paravascular cells in endosteal spaces of alveolar bone contribute to periodontal ligament cell populations. Anat Rec 219:233–242

    Article  CAS  PubMed  Google Scholar 

  • Nakashima K, de Crombrugghe B (2003) Transcriptional mechanisms in osteoblast differentiation and bone formation. Trends Genet 19:458–466

    Article  CAS  PubMed  Google Scholar 

  • Park JC, Kim JM, Jung IH, Kim JC, Choi SH, Cho KS, Kim CS (2011) Isolation and characterization of human periodontal ligament (PDL) stem cells (PDLSCs) from the inflamed PDL tissue: in vitro and in vivo evaluations. J Clin Periodontal 38:721–731

    Article  Google Scholar 

  • Pihlstrom BL, Michalowicz BS, Johnson NW (2005) Periodontal diseases. Lancet 366:1809–1820

    Article  PubMed  Google Scholar 

  • Rodríguez-Lozano FJ, Bueno C, Insausti CL, Meseguer L, Ramírez MC, Blanquer M, Marín N, Martínez S, Moraleda JM (2011) Mesenchymal stem cells derived from dental tissues. Int Endod J 44:800–806

    PubMed  Google Scholar 

  • Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, Young M, Robey PG, Wang CY, Shi S (2004) Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 364:149–155

    Article  CAS  PubMed  Google Scholar 

  • Slack JM (2000) Stem cells in epithelial tissues. Science 287:1431–1433

    Article  CAS  PubMed  Google Scholar 

  • Tomokiyo A, Maeda H, Fujii S, Wada N, Shima K, Akamine A (2008) Development of a multipotent clonal human periodontal ligament cell line. Differentiation 76:337–347

    Article  CAS  PubMed  Google Scholar 

  • Wada N, Menicanin D, Shi S, Bartold PM, Gronthos S (2009) Immunomodulatory properties of human periodontal ligament stem cells. J Cell Physiol 219:667–676

    Article  CAS  PubMed  Google Scholar 

  • Xu J, Wang W, Kapila Y, Lotz J, Kapila S (2009) Multiple differentiation capacity of STRO-1 + /CD146+ PDL mesenchymal progenitor cells. Stem Cells Dev 18:487–496

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zannettino AC, Paton S, Arthur A, Khor F, Itescu S, Gimble JM, Gronthos S (2008) Multipotential human adipose-derived stromal stem cells exhibit a perivascular phenotype in vitro and in vivo. J Cell Physiol 214:413–421

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research is funded by Vietnam Ministry of Science and Technology under grant number ĐTĐL.2012-G/34.

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Authors and Affiliations

  1. Ho Chi Minh City, Vietnam

    Ha Le Bao Tran, Vu Nguyen Doan, Huong Thi Ngoc Le & Lan Thi Quynh Ngo

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  1. Ha Le Bao Tran
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  2. Vu Nguyen Doan
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  3. Huong Thi Ngoc Le
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  4. Lan Thi Quynh Ngo
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Correspondence to Ha Le Bao Tran.

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Editor: T. Okamoto

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Tran, H.L.B., Doan, V.N., Le, H.T.N. et al. Various methods for isolation of multipotent human periodontal ligament cells for regenerative medicine. In Vitro Cell.Dev.Biol.-Animal 50, 597–602 (2014). https://doi.org/10.1007/s11626-014-9748-z

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  • DOI: https://doi.org/10.1007/s11626-014-9748-z

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