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2003 Fiscal Year Final Research Report Summary

Basic research on decelerating disc degeneration -Activation of nucleus pulposus cells-

Research Project

Project/Area Number 14571406
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Orthopaedic surgery
Research InstitutionTokai University

Principal Investigator

MOCHIDA Joji  Tokai University, School of Medicine, Professor, 医学部, 教授 (50174347)

Co-Investigator(Kenkyū-buntansha) NOMURA Takeshi  Tokai University, School of Medicine, Assistant Professor, 医学部, 講師 (60246121)
Project Period (FY) 2002 – 2003
Keywordsintervertebral disc / nucleus pulposus cells / mesenchymal stem cells / coculture
Research Abstract

NP cells were cocultured with mesenchymal stem cells (MSCs) in order to evaluate the effect of mesenchymal stem cells on activation of NP cells in vitro and in vivo. Methods : NP cells were harvested from rabbit discs. A 12-well multiple culture plates were used for coculture. A membrane culture insert was used. NP cells were seeded into each well at 1.0 X 10^4 cells / well, with or without 1.0 X 10^4 MSCs inside a membrane culture insert. On day 7 of coculture, they were evaluated for cell proliferation by using WST-8, DNA synthesis by measuring the uptake of [3H]-thymidine and proteoglycan (PG) synthesis by measuring the uptake of [35S]-sulphate. Statistical analysis by Wilcoxon's signed-ranks test was used with significance set at p<0.01. Cocultured NP cells were then reinserted into disc degeneration models in rabbits. Disc degeneration was evaluated at 2, 4, 8 and 16 weeks post reinsertion. Results : In cell proliferation assay, NP cells seeded with MSCs inside the insert (group M) was 2.97 X 10^5 cells, where as NP cells cultured monolayer (group N) only counted 4.02 X 10^4 cells / well (p<0.01). The average disintegration per second (DPM) measured for [3H]-thymidine indicating DNA synthesis was 188.56±5.20 DPM / cell in group M and 9.54±0.65 DPM / cell in group N (p<0.01). Results for [35S]-sulphate demonstrated 64.16±1.66 DPM / cell in group M and 6.42±0.15 DPM / cell in group N (p<0.01). These results correlated well with the results of in vivo studies. Conclusions : Coculture of nucleus pulposus cells by mesenchymal stem cells resulted in an improvement of cell proliferation and matrix synthesis over monolayer culture and reinsertion of nucleus pulposus cells activated by mesenchymal stem cells using coculture method was more efficient than our previous studies in deceleration of intervertebral disc degeneration.

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] 山本至宏, 持田譲治ほか: "椎間板変性抑制のための髄核再挿入術 再挿入髄核活性化の検討"日本脊椎脊髄病学会雑誌. 13巻1号. 276 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 酒井大輔, 持田譲治ほか: "髄核細胞の再挿入による椎間板変性抑制機序について-その効率向上への骨髄間葉系幹細胞の関与-"日本整形外科学会雑誌. 76巻8号. S905 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本至宏, 持田譲治ほか: "椎間板変性抑制のための基礎的研究-骨髄間葉系幹細胞とのCocultureによる同種髄核細胞の再挿入術-"日本整形外科学会雑誌. 76巻8号. S987 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本至宏, 持田譲治ほか: "骨髄間葉系幹細胞による同種髄核細胞の活性化 Coculture法における細胞間接着の意義"日本脊椎脊髄病学会雑誌. 14巻1号. 121 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本至宏, 持田譲治ほか: "骨髄間葉系幹細胞と髄核細胞のCocultureが間葉系幹細胞の分化に与える影響"日本整形外科学会雑誌. 77巻8号. S1067 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 酒井大輔, 持田譲治ほか: "幹細胞を用いた細胞移植による椎間板再生"日本整形外科学会雑誌. 77巻8号. S925 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Watanabe K, Mochida J et al.: "Effect of reinsertion of activated nucleus pulposus on disc degeneration : an experimental study on various types of collagen in degenerative discs"Connect Tissue Res.. 44(2). 104-108 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Reinsertion of nucleus pulposus cells activated by mesenchymal stem cells using coculture method decelerated intervertebral disc degeneration."Spine Journal. 3(5S). 101S (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Upregulation of viability of nucleus pulposus cells using novel coculture system -significance of direct cellular interaction-."Spine. 29(in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Activated nucleus pulposus reinsertion for the treatment of disc degeneration."The Journal of the Japan Spine Research Society. 13-1. 276 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sakai D, Mochida J et al.: "Deceleration of intervertebral disc by reinsertion of nucleus pulposus cells."Nihon Seikei Geka Gakkaishi. 76-8. S905 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Basic research for the decleration of disc degeneration"Nihon Seikei Geka Gakkaishi. 76-8. S987 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Upregulation of viability of nucleus pulposus cells using novel coculture system -significance of direct cellular interaction-."The Journal of the Japan Spine Research Society. 14-1. 121 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Effect of MSC-NP coculture on differentiation of MSCs."Nihon Seikei Geka Gakkaishi. 77-8. S1067 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sakai D, Mochida J et al.: "Stem cell applications in regeration of intervertebral disc."Nihon Seikei Geka Gakkaishi. 77-8. S925 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe K, Mochida J et al.: "Effect of reinsertion of activated nucleus pulposus on disc degeneration : an experimental study on various types of collagen in degenerative discs."Connect Tissue Res.. 44-2. 104-108 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Reinsertion of nucleus pulposus cells activated by mesenchymal stem cells using coculture method decelerated intervertebral disc degeneration."Spine Journal. 3(5S). 101S (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto Y, Mochida J et al.: "Upregulation of viability of nucleus pulposus cells using novel coculture system -significance of direct cellular interaction-."Spine. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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