| Project/Area Number |
10357015
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Ophthalmology
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| Research Institution | Chiba University |
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Principal Investigator |
ADACHI Emiko Department of Ophthalmology, Chiba University School of Medicine, Professor, 医学部, 教授 (60009496)
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| Co-Investigator(Kenkyū-buntansha) |
TSUYAMA Yoshihiko Department of Ophthalmology, Chiba University School of Medicine, Associate Professor, 医学部・附属病院, 講師 (50210571)
MIZOTA Atsushi Department of Ophthalmology, Chiba University School of Medicine, Assistant Professor, 医学部, 講師 (10239262)
FUJIMOTO Naoya Department of Ophthalmology, Chiba University School of Medicine, Associate Professor, 医学部, 助教授 (10199375)
出沢 真理 千葉大学, 医学部, 助手 (50272323)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥32,500,000 (Direct Cost: ¥32,500,000)
Fiscal Year 2000: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1999: ¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1998: ¥18,900,000 (Direct Cost: ¥18,900,000)
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| Keywords | optic nerve / gene transfer / neurotrophic factor / superior colliculus / transplantation / 架橋手術 / 細胞外基質 / 再生 / シュワン細胞 |
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| Research Abstract |
We investigate whether the graft made by cultured Schwann cell can provide a substrate enabling regrowth of injured optic nerve, and assess the effect of neurotrophic factors for the induction of optic nerve regeneration. Cultured Schwann cells were purified from dorsal root ganglions of newborn rats (Wistar strain). Approximately 106 Schwann cells/ml were suspended in an extracellular matrix containing neurotrophic factors either of NGF, BDNF or NT4, and then transferred into a silicon tube or a hollow fiber. The grafts were transplanted to the retinal stump of the transected adult optic nerve (Wistar strain). The degree of optic nerve regeneration was evaluated by counting the number of diI labeled retinal ganglion cells (RGCs). The degree of surviving RGCs was 2±1.01 % only with the extracellular matrix. The combination with BDNF or CNTF intravitreous injection resulted in an increase of 30.28±16.97 % or 54.32±8.005 %. As the second step, we elicited the regrowth of transected optic nerve fibers into superior colliculus by this artificial graft. The grafts linked the left eye and the right superior colliculus along a extracranial course in adult rats (Wistar strain). Three months after operation, diI retrograde labeling (injected into the superior colliculus) showed that 18.1±0.11 % of retinal ganglion cell axons penetrated the superior colliculus. Furthermore, we evaluated the in vivo electroporation as a delivery method in RGCs, efficiency, cell specificity, stability and tissue damage were investigated by using green fluorescent protein gene as the genetic marker. The results showed that approximately 40 % of total ganglion cells in the entire electroinjected area were GFP positive by 7th day.
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