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2006 Fiscal Year Final Research Report Summary

Dentin regeneration using pulp cells fused with immunodeficient amniotic stem cells

Research Project

Project/Area Number 17390509
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Conservative dentistry
Research InstitutionNational Institute for Longevity Sciences, National Center for Geriatrics and Gerontology

Principal Investigator

NAKASHIMA Misako  National Institute for Longevity Sciences, Lab.of Oral Disease Research, Chief, (研究所)・口腔疾患研究部, 室長 (20207773)

Co-Investigator(Kenkyū-buntansha) MATSUSHIMA Kenji  National Institute for Longevity Sciences, Lab.of Oral Disease Research, Head, (研究所)・口腔疾患研究部, 部長 (90253898)
HASHIMOTO Naohiro  National Institute for Longevity Sciences, Dept.of Regenerative Medicine, Head, (研究所)・再生再建医学研究部, 部長 (00208456)
Project Period (FY) 2005 – 2006
Keywordspulp stem cells / amniotic stem cells / dentin regeneration / Side population (SP) cells / flow cytometry / laser microdissection / immunodeficiency / vasculogenesis / angiogenesis
Research Abstract

The primary amniotic cells from mouse E14 embryo were separated by digestion with dispase and collagenase. CD44^+ ; Sca1^+ cells were further isolated by flow cytometry and expanded on feeders. Those CD44^+; Sca1^+ cells strongly expressed embryonic and adult stem cell markers, POU/Oct4, Sox2, Tert, Rex1, nanog. We had previously isolated side population (SP) cells from porcine dental pulp, which were enriched for stem cell properties. The mRNA expression of the endothelial cell markers, CD31, CD105, Vgfr2 were much stronger in SP cells compared with non-SP cells. To isolate further distinct populations, flow cytometric analyses of the SP cells were performed using antibodies against CD31 and CD146. The CD31^-population was almost CD146^-and represented 50% of total SP cells. The CD31^+ populatio contained both CD146^-and CD146^+ cells, representing 2% and 48% of total SP cells, respectively. CD31^- ; CD146^-SP cells have high vasculogenic potential as well as multi-differentiation potential including odontogenic potential, highly proliferative and migration activities. The subtraction microarray analysis and laser microdissection analysis showed that thyrotropin-releasing hormone degrading ectoenzyme (TRHDE) and low density lipoprotein-related protein 1B (LRP1B) were expressed highly in the pulp stem cells but not in the amniotic stem cells, suggesting possible markers for transdifferentiation of amniotic stem cells. We tried to transdifferentiate the amniotic stem cells into the odontoblastic lineage by some gene electrotransfection, analyzing by expression of TRHDE and differentiation markers for odontoblasts.

  • Research Products

    (23 results)

All 2007 2006 2005

All Journal Article (21 results) Book (2 results)

  • [Journal Article] Runx3 negatively regulates Osterix expression in dental pulp cells.2007

    • Author(s)
      Zheng L, Iohara K, Ishikawa M, Into T, Takano-Yamamoto T, Matsushita K, Nakashima M
    • Journal Title

      Biochem J. Mar 13(in press)

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Pathogen recognition by Toll-like receptor 2 activates Weibel-Palade body exocytosis in human aortic endothelial cells.2007

    • Author(s)
      Into T, Kanno Y, Dohkan J, Nakashima M, Inomata M, Shibata K, Lowenstein CJ, Matsushita K
    • Journal Title

      J Biol Chem. 282・11

      Pages: 8134-8141

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Synthesis and Characterization of a Dipalmitoylated Lipopeptide Derived from Paralogous Lipoproteins of Mycoplasma pneumoniae.2007

    • Author(s)
      Into T, Dohkan J, Inomata M, Nakashima M, Shibata K, Matsushita K
    • Journal Title

      Infect Immun. 75・5

      Pages: 2253-2259

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Runx3 negatively regulates Osterix expression in dental pulp cells.2007

    • Author(s)
      Zheng L, Iohara K, Ishikawa M, Into T, Takano-Yamamoto T, Matsushita K, Nakashima M.
    • Journal Title

      Biochem J (in press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Synthesis and Characterization of a Dipalmitoylated Lipopeptide Derived from Paralogous Lipoproteins of Mycoplasma pneumoniae.2007

    • Author(s)
      Into T, Dohkan J, Inomata M, Nakashima M, Shibata K, Matsushita K.
    • Journal Title

      J Biol Chem 282-11

      Pages: 8134-8141

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Tissue engineering in endodontics.2007

    • Author(s)
      Nakashima M.
    • Journal Title

      Infect Immun. 75-5

      Pages: 2253-2259

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Tissue Engineering of Tooth.2007

    • Author(s)
      Nakashima M.
    • Journal Title

      Handbook of Biomineralization. (Wiley-VCH publisher)

      Pages: 265-281

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] The side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis and neurogenesis.2006

    • Author(s)
      Iohara K, Zheng L, Ito M, Tomokiyo A, Matsushita K, Nakashima M
    • Journal Title

      Stem Cells 24・11

      Pages: 2493-2503

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Gene therapy for dentin regeneration with bone morphogenetic proteins.2006

    • Author(s)
      Nakashima M, Iohara K, Zheng L
    • Journal Title

      Current Gene Ther. 6・5

      Pages: 551-560

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Arginine-specific gingipains from Porphyromonas gingivalis deprive protective functions of secretory leucocyte protease inhibitor in periodontal tissue.2006

    • Author(s)
      Into T, Inomata M, Kanno Y, Matsuyama T, Machigashira M, Izumi Y, Imamura T, Nakashima M, Noguchi T, Matsushita K
    • Journal Title

      Clin Exp Immunol. 145・3

      Pages: 545-554

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis and neurogenesis.2006

    • Author(s)
      Iohara K, Zheng L, Ito M, Tomokiyo A, Matsushita K, Nakashima M.
    • Journal Title

      Stem Cells 24-11

      Pages: 2493-2503

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Gene therapy for dentin regeneration with bone morphogenetic proteins. Arginine-specific gingipains from Porphyromonas gingivalis deprive protective functions of secretory leucocyte protease inhibitor in periodontal tissue.2006

    • Author(s)
      Nakashima M, Iohara K, Zheng L
    • Journal Title

      Current Gene Then 6-5

      Pages: 551-560

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Pathogen recognition by Toll-like receptor 2 activates Weibel-Palade body exocytosis in human aortic endothelial cells.2006

    • Author(s)
      Into T, Inomata M, Kanno Y, Matsuyama T, Machigashira M, Izumi Y, Imamura T, Nakashima M, Noguchi T, Matsushita K. Into T, Kanno Y, Dohkan J, Nakashima M, Inomata M, Shibata K, Lowenstein CJ, Matsushita K.
    • Journal Title

      Clin Exp Immunol. 145-3

      Pages: 545-554

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] 象牙芽細胞分化過程における転写因子Osterixの発現とその調節機構2005

    • Author(s)
      石河真幸, 中島美砂子, 庵原耕一郎, 赤峰昭文
    • Journal Title

      日本歯科保存学雑誌 48・6

      Pages: 813-821

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Tissue engineering in endodontics.2005

    • Author(s)
      Nakashima M
    • Journal Title

      Aust Endod J. 31・3

      Pages: 111-113

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Growth/differentiation factor 11 (Gdf11)を電気的遺伝子導入した歯髄幹細胞を用いた生体外遺伝子導入法による修復象牙質形成2005

    • Author(s)
      中島美砂子
    • Journal Title

      日本歯科保存学雑誌. 48・6

      Pages: 803-805

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The application of tissue engineering to regeneration of pulp and dentin in endodontics.2005

    • Author(s)
      Nakashima M., Akamine A
    • Journal Title

      J. Endodontics 31・10

      Pages: 711-718

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Bone morphogenetic proteins in dentin regeneration for potential use in endodontic therapy.2005

    • Author(s)
      Nakashima M
    • Journal Title

      Cytokine Growth Factor Rev 16・3

      Pages: 369-376

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The application of tissue engineering to regeneration of pulp and dentin in endodontics.2005

    • Author(s)
      Nakashima M, Akamine A.
    • Journal Title

      Aust Endod J. 31-11

      Pages: 111-113

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Bone morphogenetic proteins in dentin regeneration for potential use in endodontic therapy.2005

    • Author(s)
      Nakashima M.
    • Journal Title

      J. Endodontics 31-10

      Pages: 711-718

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Tissue Engineering of Tooth.2005

    • Author(s)
      Nakashima M.
    • Journal Title

      Cytokine Growth Factor Rev 16-3

      Pages: 369-376

    • Description
      「研究成果報告書概要(欧文)」より
  • [Book] Handbook of Biomineralization. Tissue Engineering of Tooth. Vol. 3. Chapter 172007

    • Author(s)
      Nakashima M
    • Total Pages
      265-281(17)
    • Publisher
      Wiley-VCH publisher
    • Description
      「研究成果報告書概要(和文)」より
  • [Book] 歯の再生 歯の発生生物学から歯の再生研究まで 第4章歯の再生 7.歯髄幹細胞を用いた象牙質・歯髄再生2006

    • Author(s)
      中島美砂子
    • Total Pages
      192-199(8)
    • Publisher
      真興交易(株)医書出版部
    • Description
      「研究成果報告書概要(和文)」より

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Published: 2008-05-27  

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