| Project/Area Number |
63570688
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | The Jikei University School of Medicine |
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Principal Investigator |
TANI Satoshi (1989-1990) The Jikei Univ. School of Medicine, Neurosurgery Post Doctorfellow-Ship, 医学部・脳神経外科, 助手 (10147332)
篠田 宗次 (1988) 東京慈恵会医科大学, 医学部, 講師 (80110922)
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| Co-Investigator(Kenkyū-buntansha) |
SHINODA Souji Jikei Medical School of Medicine, Neurosurgery Associate Proofessor, 脳神経外科, 助教授 (80110922)
NAKAMURA Norio The Jikei Univ. School of Medicine, Neurosurgery Professor, 脳神経外科, 教授 (30056494)
谷 諭 東京慈恵会医科大学, 医学部, 助手 (10147332)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1988: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Central Neuron / Tissue Culture / Cell Culture / Neurofilament / Neuron Growth Factor / 中枢神経培養 / 成長因子 / Neurofilament / 68K / 160k / 200k / 鶏中枢神経 / 培養 |
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| Research Abstract |
Primary neuron cells of check embryonic brain were cultured and these neuron cells were shown good net-work relation of each others. These neuron cell bodies, dendrites and axons were demonstrated with histochemical technique of Neurofilaments (NF), neuron specific enolase and tetanus toxin which were related with special identify to neuron cells. (1) Appearance of each neurons network were better from tissue culture of central neuron method than cell suspension culture with 100 mum mesh, although we could see more beautiful one neuron cell shape from cell suspension than tissue culture method. (2) Identification of neuron cell were stained histochemically by avidin-biotin complex technique using 68K, 160K and whole NF human anti-serum which were purchased from Cosmo bio. But cross reaction of chick neuron cells and anti human NF serum had to be certificated. These reactions were demonstrated by western-blotting technique which protein components were separated by SDS polyacrylamide ge
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l electrophoresis and showing at the positions of 68K and 160K dalton. These protein components were got by pure neuron cells using poly-L-lysine culture. (3) The results of histochemical staining of 68K and 160K NF were different in chick neuron cells. 68K NF existed at the cell body, axon and dendrite, although 160K NF existed at the cell body but few at the axon and dendrite. Such a facts were speculated that slow transportation of NF on the axon and dendrite is related only 68K and not 68K and suspected of changing molecular weight 160K NF to 68K NF when something is need to transport of NF. (4) Alkaline-phosphatese and protease enzyme which digest of 160K NF and dephosphrylate of 200K NF were treated to the cultured neuron cells, because we tried to certification of NF transportation on the axon and dendrite. These enzymes were used for 3-12 hours at 6-8 days neuron cells after primary culture. But there were no difference of histochemical appearance between treated these enzymes and control. But we should try again and use another technique or new method. (5) Although neurons growth and network in the neuron culture with temporal lobe were better impression of growth than without temporal lobe, cerebellar neurons culture were no difference with and without temporal lobe. And neuron culture with and without temporal components were not related of long survival of neuron cells life in vitro. These research should be done continuously to get long survival central neuron cells. This theme is important and will be related to save many patients who are now painful of hemiparesis, aphasia, visual field disturbance and others. Less
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