| Project/Area Number |
06454361
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | UNIVERSITY OF TOKYO |
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Principal Investigator |
MIYATA Tetsuro SECOND DEPARTMENT OF SURGERY,UNIVERSITY OF TOKYO,ASSOCIATE IN SURGERY, 医学部・附属病院, 助手 (70190791)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Osamu SECOND DEPARTMENT OF SURGERY,UNIVERSITY OF TOKYO,ASSOCIATE IN SURGERY, 医学部・附属病院, 助手 (40170724)
IWAMOTO Aikichi NATIONAL INSTITUTE OF HEALTH AND INSTITUTE OF MEDICAL SCIENCE,UNIVERSITY OF TOKY, 医科学・研究所, 教授 (10133076)
小山 博之 東京大学, 医学部(病), 助手 (10241994)
木村 秀生 東京大学, 医学部(病), 助手
高木 淳彦 東京大学, 医学部(病), 講師 (50092979)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1996: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1995: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1994: ¥3,700,000 (Direct Cost: ¥3,700,000)
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| Keywords | gene therapy / adenovirus / tissue plasminogen activator / thrombomodulin / hybrid-type vascular prosthesis / endothelial cell / small caliber vascular prosthesis / retrovirus / vetrovirus |
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| Research Abstract |
Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secretine anti-thrombotic agents has considerable clinical potential. Adenoviral vactor was used to transfer human thrombomodulin (hTM) or human tissue plasminogen activator (htPA) genes to hECs and the ability of the transduced hECs was examined. Cultured hECs on plates were infected with various concentrations of recombinant adenoviruses containing the hTM or htPA gene to determine the appropriate transfer conditions. Furthermore, htPA gene transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4mm internal diameter), some of which were exposed to pulsatile flow in vitro to check the ability of secreting htPA in the circuit, and retention of the hECs on the graft. Effective transduction of hTM and htPA gene into hECs was achieved with viral soup at 30 pfu/ml : hTM antigen 589+-42.3 MGCC U/mg protein, hTM protein C-activating cofactor activity 32.4+-10.1 U/ml, htPA antigen 4720+-86 ng/106 cells/6h, htPA activity 647+-32 IU/106 cells/6h. Grafts seeded with htPA gene transduced hECs retained the satisfactory secretion of htPA in the in vitro flow circuit. The retention of the perfused grafts with tPA transduced ECs was 84.0+-3.0%, with mock-infected (LacZ) ECs 94.2+-0.4%, and with the non-infected ECs 82.1+-8.0%. There were no significant differences among these retention values. There results indicate that seeding small-caliber vascular grafts with genetically engineered hECs has the potential to increase local anti-thrombotic properites in vivo.
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