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2003 Fiscal Year Final Research Report Summary

Experimental studies on peripheral nerve regeneration through the bio-nerve with 3D cultured Schwann cell

Research Project

Project/Area Number 14571401
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Orthopaedic surgery
Research InstitutionKEIO UNIVERSITY

Principal Investigator

NAKAO Yasushi  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (30188883)

Co-Investigator(Kenkyū-buntansha) NISHIWAKI Masao  KEIO UNIV., School of Medicine, Instructor, 医学部, 助手 (90296640)
TAKAYAMA Shinichiro  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (40138045)
HIROTAKA James  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (90338020)
MORISAWA Yasushi  KEIO UNIV., School of Medicine, Instructor, 医学部, 助手 (60306751)
Project Period (FY) 2002 – 2003
KeywordsSchwann cell / artificial nerve / nerve graft
Research Abstract

Schwann cells were initially isolated P7 Wister rat sciatic nerve and dorsal root ganglia. The neurotrophic factor cDNA was inserted into multiple cloning site of shuttle vector. The adenoviral constructs were transfected into 293 packaging cell line. The cultured Schwann cells were infected with concentrated adenoviruses. The expression of the target neurotrophic factor gene was evaluated by Western blot analysis. Neurons from rat DRGs were cultured in a medium obtained from the transferred Schwann cell culture at 3days, 3 weeks, 8 weeks after infection. Neurite outgrowth was measured by immunostaining of axons with monoclonal antibody RT97. A 20mm nerve gap was bridged with a bioabsorbable polymer conduit containing a mixture of the transferred Schwann cells and Pluronic gel. 12 weeks later, histological evaluation was performed at the midportion of the grafted lesion. The transferred Schwann cells lysates showed the higher expression of the transgene product of 30 Kilo Dalton level by Western blot analysis. Histological evaluation demonstrated that, contrary to the poor regeneration in the control group, the genetically modified group showed not only well-regenerated axons, but also formations of vessels and epineurium-like tissue observed.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Komiyama T: "A novel technique for peripheral nerve reconstruction in the absence of an artificial conduit."J.Neuroscience Methods. 134. 133-140 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Komiyama T: "A novel technique to isolate adult Schwann cells for an artificial nerve conduit."J.Neuroscience Methods. 122. 195-200 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 栩木弘和: "シュワン細胞の3次元培養を応用した人工神経の開発"整形災害外科. 46. 247-251 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 仲尾保志: "ティッシュエンジニアリングを応用した人工神経の開発"末梢神経. 14. 7-16 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Komiyama T, Nakao Y, Toyama Y, Vacanti CA, Vacanti MP, Ronald Ignotz: "A novel technique for peripheral nerve reconstruction in the absence of an artificial conduit."Journal of Neuroscience Methods. 139. 133-140 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Komiyama T, Nakao Y, Toyama Y, Vacanti CA, Vacanti MP, Asou H.: "A novel technique to isolate adult Schwann cells for an artificial nerve conduit."Journal of Neuroscience Methods. 122. 195-200 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tochigi N, Nakao Y: "Artificial peripheral nerve using 3D culturing of Schwann cells."Seikei-saigaigeka. 46. 247-251 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakao Y, Ikada Y: "Tissue-engineered peripheral nerve."Peripheral Nerve. 14. 7-16 (2003)

    • 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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