| Project/Area Number |
62570368
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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 |
Neurology
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| Research Institution | Department of Neurology, Nara Medical University |
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Principal Investigator |
TAKAYANAGI TETSUY Department of Neurology,Nara Medical University,Professor, 医学部・神経内科, 教授 (60022836)
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| Co-Investigator(Kenkyū-buntansha) |
堀川 博誠 奈良県立医科大学, 医学部・神経内科, 助手 (20192320)
FUMAKAWA ITARU Department of Neurology,Nara Medical University,Instructor, 医学部・神経内科, 助手 (30144819)
HORIKAWA HIROSE Department of Neurology,Nara Medical University,Instructor, 医学部・神経内科, 講師 (00145880)
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| Project Period (FY) |
1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | 血管形成因子 / HT-29 / 成長因子 |
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| Research Abstract |
Angiogenin,which was isolated and characterized by Fett,J.W. et al in 1985 and amino acid sequence was determined,is a kind of growth factor to play a role of tissue reaction. We performed to isolate angiogenin from HT-29,being human colon adenocarcinoma cells, by the method as described by Fett,J.W. et al.
Large amount of HT-29 cells was cultured in cell factory utilizing the medium of DMEM containing 5% fetal calf serum. Thereafter the cultured cells were maintained in serum-free medium,which was changed every 2-3 days. The collected serum-free media were acidified by acetate and was treated with Pepstation A. The filtrate was subsequently concentrated 200-fold, dialyzed and lyophilized. Its solution was chromatographed on CM cellulose and 1 M NaCl eluted fractions were dialyzed and lyophilized. 35mg. of solid material was obtained from 8000 ml.of serum-free medium of HT-29. Reversed-phase HPLC was performed to isolate angiogenin from the material reconstituted in 0.1% TFA. Columns were eluted with linear gradients using the same method as described by Fett,J@.w. et al. However,the amount of angiogenin showed 0.3 g. in 100 ml. of serum-free medium of HT-29. The very small quantity of angiogenin was not enough to make further studies.We are now endeavouring to have a stock of the material to proceed to immunological works.
Angiogenin preparation is being tried from other tissues,which contain more amount of angiogenin as compared with tumor. Angiogenin was clarified recently to exist in a large amount in other tissues than tumor,which let us conduct a research in different ways.
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