2003 Fiscal Year Final Research Report Summary
Experimental studies on peripheral nerve regeneration through the bio-nerve with 3D cultured Schwann cell
| Project/Area Number |
14571401
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | KEIO UNIVERSITY |
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Principal Investigator |
NAKAO Yasushi KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (30188883)
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| 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)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Schwann cell / artificial nerve / nerve graft |
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| 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.
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