Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells
Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secreting antithrombogenic or fibrinolytic agents has considerable clinical potential. An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of t...
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| Veröffentlicht in: | Surgery 1997-07, Vol.122 (1), p.91-100 |
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| creator | SUGAWARA, Y SAKATA, Y MIYATA, T MINOWADA, S HAMADA, H YOSHIDA, Y SATO, O DEGUCHI, J.-O KIMURA, H NAMBA, T MAKUUCHI, M |
| description | Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secreting antithrombogenic or fibrinolytic agents has considerable clinical potential.
An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of the transduced hECs to secrete htPA was examined. Cultured hECs on plates were incubated with various concentrations of recombinant adenoviruses containing the htPA or LacZ gene for various times to determine the optimal transfer conditions. Transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4 mm in diameter), some of which were exposed to pulsatile flow in vitro.
Effective transduction of the htPA gene into hECs (htPAhECs) was achieved with viral soup at a multiplicity of infection of 30 after incubation for 1 day, which yielded 4.8 +/- 0.20 x 10(3) ng/10(6) cells/6 hr htPA antigen on plates (n = 3), 2.2 +/- 2.0 x 10(3) ng/10(6) cells/6 hr on grafts (n = 6), and 6.8 +/- 1.7 x 10(2) ng/10(6) cells/6 hr on perfused grafts (n = 6). The retention of htPAhECs by perfused grafts was 84.0% +/- 3.0%, comparable with the noninfected (82.1% +/- 8.0%) and mock-infected (94.2% +/- 0.4%) hEC values.
By adenoviral vector-mediated gene transfer, 10(2-3)-fold enhancement of htPA secretion was demonstrated, which did not affect cell retention by grafts. |
| doi_str_mv | 10.1016/S0039-6060(97)90269-5 |
| format | Article |
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An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of the transduced hECs to secrete htPA was examined. Cultured hECs on plates were incubated with various concentrations of recombinant adenoviruses containing the htPA or LacZ gene for various times to determine the optimal transfer conditions. Transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4 mm in diameter), some of which were exposed to pulsatile flow in vitro.
Effective transduction of the htPA gene into hECs (htPAhECs) was achieved with viral soup at a multiplicity of infection of 30 after incubation for 1 day, which yielded 4.8 +/- 0.20 x 10(3) ng/10(6) cells/6 hr htPA antigen on plates (n = 3), 2.2 +/- 2.0 x 10(3) ng/10(6) cells/6 hr on grafts (n = 6), and 6.8 +/- 1.7 x 10(2) ng/10(6) cells/6 hr on perfused grafts (n = 6). The retention of htPAhECs by perfused grafts was 84.0% +/- 3.0%, comparable with the noninfected (82.1% +/- 8.0%) and mock-infected (94.2% +/- 0.4%) hEC values.
By adenoviral vector-mediated gene transfer, 10(2-3)-fold enhancement of htPA secretion was demonstrated, which did not affect cell retention by grafts.</description><identifier>ISSN: 0039-6060</identifier><identifier>EISSN: 1532-7361</identifier><identifier>DOI: 10.1016/S0039-6060(97)90269-5</identifier><identifier>PMID: 9225920</identifier><identifier>CODEN: SURGAZ</identifier><language>eng</language><publisher>New York, NY: Elsevier</publisher><subject>Adenoviridae - genetics ; Adult ; Base Sequence ; Biological and medical sciences ; Cell Count ; Cells, Cultured ; Electrophoresis, Polyacrylamide Gel ; Endothelium, Vascular - chemistry ; Endothelium, Vascular - cytology ; Endothelium, Vascular - metabolism ; Fibrin - analysis ; Gene Expression - physiology ; Gene Transfer Techniques - standards ; Genetic Vectors ; Humans ; Immunoblotting ; Medical sciences ; Miscellaneous ; Polytetrafluoroethylene ; Prostheses and Implants ; Pulsatile Flow ; Saphenous Vein - cytology ; Sodium Dodecyl Sulfate ; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases ; Tissue Plasminogen Activator - analysis ; Tissue Plasminogen Activator - genetics ; Tissue Plasminogen Activator - metabolism</subject><ispartof>Surgery, 1997-07, Vol.122 (1), p.91-100</ispartof><rights>1997 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2746188$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9225920$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>SUGAWARA, Y</creatorcontrib><creatorcontrib>SAKATA, Y</creatorcontrib><creatorcontrib>MIYATA, T</creatorcontrib><creatorcontrib>MINOWADA, S</creatorcontrib><creatorcontrib>HAMADA, H</creatorcontrib><creatorcontrib>YOSHIDA, Y</creatorcontrib><creatorcontrib>SATO, O</creatorcontrib><creatorcontrib>DEGUCHI, J.-O</creatorcontrib><creatorcontrib>KIMURA, H</creatorcontrib><creatorcontrib>NAMBA, T</creatorcontrib><creatorcontrib>MAKUUCHI, M</creatorcontrib><title>Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells</title><title>Surgery</title><addtitle>Surgery</addtitle><description>Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secreting antithrombogenic or fibrinolytic agents has considerable clinical potential.
An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of the transduced hECs to secrete htPA was examined. Cultured hECs on plates were incubated with various concentrations of recombinant adenoviruses containing the htPA or LacZ gene for various times to determine the optimal transfer conditions. Transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4 mm in diameter), some of which were exposed to pulsatile flow in vitro.
Effective transduction of the htPA gene into hECs (htPAhECs) was achieved with viral soup at a multiplicity of infection of 30 after incubation for 1 day, which yielded 4.8 +/- 0.20 x 10(3) ng/10(6) cells/6 hr htPA antigen on plates (n = 3), 2.2 +/- 2.0 x 10(3) ng/10(6) cells/6 hr on grafts (n = 6), and 6.8 +/- 1.7 x 10(2) ng/10(6) cells/6 hr on perfused grafts (n = 6). The retention of htPAhECs by perfused grafts was 84.0% +/- 3.0%, comparable with the noninfected (82.1% +/- 8.0%) and mock-infected (94.2% +/- 0.4%) hEC values.
By adenoviral vector-mediated gene transfer, 10(2-3)-fold enhancement of htPA secretion was demonstrated, which did not affect cell retention by grafts.</description><subject>Adenoviridae - genetics</subject><subject>Adult</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cell Count</subject><subject>Cells, Cultured</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Endothelium, Vascular - chemistry</subject><subject>Endothelium, Vascular - cytology</subject><subject>Endothelium, Vascular - metabolism</subject><subject>Fibrin - analysis</subject><subject>Gene Expression - physiology</subject><subject>Gene Transfer Techniques - standards</subject><subject>Genetic Vectors</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Medical sciences</subject><subject>Miscellaneous</subject><subject>Polytetrafluoroethylene</subject><subject>Prostheses and Implants</subject><subject>Pulsatile Flow</subject><subject>Saphenous Vein - cytology</subject><subject>Sodium Dodecyl Sulfate</subject><subject>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</subject><subject>Tissue Plasminogen Activator - analysis</subject><subject>Tissue Plasminogen Activator - genetics</subject><subject>Tissue Plasminogen Activator - metabolism</subject><issn>0039-6060</issn><issn>1532-7361</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kEtLxDAUhYMo4zj6EwayENFF9SZp0mY5iC8QXKjr4ba9cSJ92aQD8--tOMzqcjgfh3MPY0sBtwKEuXsHUDYxYODaZjcWpLGJPmJzoZVMMmXEMZsfkFN2FsI3ANhU5DM2s1JqK2HOcFVR2239MIakocpjpIrHAdvgaOCd49GHMFISdz3xvsbQ-Lb7opZjGf0WYzfwSRGPHd-MDbac2qqLG6o91rykug7n7MRhHehifxfs8_Hh4_45eX17erlfvSa9VFlM0GirUkyFE4KqvKigkFpnJjfOggPIAbUsZYE6zQpJqcqcJOtQCkfKaacW7Oo_tx-6n5FCXDc-_DXAlroxrDMrlFU5TOByD47F9PK6H3yDw26932TyL_c-hhJrN41R-nDAZJYakefqF7TocqE</recordid><startdate>19970701</startdate><enddate>19970701</enddate><creator>SUGAWARA, Y</creator><creator>SAKATA, Y</creator><creator>MIYATA, T</creator><creator>MINOWADA, S</creator><creator>HAMADA, H</creator><creator>YOSHIDA, Y</creator><creator>SATO, O</creator><creator>DEGUCHI, J.-O</creator><creator>KIMURA, H</creator><creator>NAMBA, T</creator><creator>MAKUUCHI, M</creator><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>19970701</creationdate><title>Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells</title><author>SUGAWARA, Y ; SAKATA, Y ; MIYATA, T ; MINOWADA, S ; HAMADA, H ; YOSHIDA, Y ; SATO, O ; DEGUCHI, J.-O ; KIMURA, H ; NAMBA, T ; MAKUUCHI, M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p237t-a65934a41f11ed8bd0b2557686f90f0080a52c2ba547b2e437f2e9fa21fe3f5f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Adenoviridae - genetics</topic><topic>Adult</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cell Count</topic><topic>Cells, Cultured</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Endothelium, Vascular - chemistry</topic><topic>Endothelium, Vascular - cytology</topic><topic>Endothelium, Vascular - metabolism</topic><topic>Fibrin - analysis</topic><topic>Gene Expression - physiology</topic><topic>Gene Transfer Techniques - standards</topic><topic>Genetic Vectors</topic><topic>Humans</topic><topic>Immunoblotting</topic><topic>Medical sciences</topic><topic>Miscellaneous</topic><topic>Polytetrafluoroethylene</topic><topic>Prostheses and Implants</topic><topic>Pulsatile Flow</topic><topic>Saphenous Vein - cytology</topic><topic>Sodium Dodecyl Sulfate</topic><topic>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</topic><topic>Tissue Plasminogen Activator - analysis</topic><topic>Tissue Plasminogen Activator - genetics</topic><topic>Tissue Plasminogen Activator - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>SUGAWARA, Y</creatorcontrib><creatorcontrib>SAKATA, Y</creatorcontrib><creatorcontrib>MIYATA, T</creatorcontrib><creatorcontrib>MINOWADA, S</creatorcontrib><creatorcontrib>HAMADA, H</creatorcontrib><creatorcontrib>YOSHIDA, Y</creatorcontrib><creatorcontrib>SATO, O</creatorcontrib><creatorcontrib>DEGUCHI, J.-O</creatorcontrib><creatorcontrib>KIMURA, H</creatorcontrib><creatorcontrib>NAMBA, T</creatorcontrib><creatorcontrib>MAKUUCHI, M</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>SUGAWARA, Y</au><au>SAKATA, Y</au><au>MIYATA, T</au><au>MINOWADA, S</au><au>HAMADA, H</au><au>YOSHIDA, Y</au><au>SATO, O</au><au>DEGUCHI, J.-O</au><au>KIMURA, H</au><au>NAMBA, T</au><au>MAKUUCHI, M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells</atitle><jtitle>Surgery</jtitle><addtitle>Surgery</addtitle><date>1997-07-01</date><risdate>1997</risdate><volume>122</volume><issue>1</issue><spage>91</spage><epage>100</epage><pages>91-100</pages><issn>0039-6060</issn><eissn>1532-7361</eissn><coden>SURGAZ</coden><abstract>Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secreting antithrombogenic or fibrinolytic agents has considerable clinical potential.
An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of the transduced hECs to secrete htPA was examined. Cultured hECs on plates were incubated with various concentrations of recombinant adenoviruses containing the htPA or LacZ gene for various times to determine the optimal transfer conditions. Transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4 mm in diameter), some of which were exposed to pulsatile flow in vitro.
Effective transduction of the htPA gene into hECs (htPAhECs) was achieved with viral soup at a multiplicity of infection of 30 after incubation for 1 day, which yielded 4.8 +/- 0.20 x 10(3) ng/10(6) cells/6 hr htPA antigen on plates (n = 3), 2.2 +/- 2.0 x 10(3) ng/10(6) cells/6 hr on grafts (n = 6), and 6.8 +/- 1.7 x 10(2) ng/10(6) cells/6 hr on perfused grafts (n = 6). The retention of htPAhECs by perfused grafts was 84.0% +/- 3.0%, comparable with the noninfected (82.1% +/- 8.0%) and mock-infected (94.2% +/- 0.4%) hEC values.
By adenoviral vector-mediated gene transfer, 10(2-3)-fold enhancement of htPA secretion was demonstrated, which did not affect cell retention by grafts.</abstract><cop>New York, NY</cop><pub>Elsevier</pub><pmid>9225920</pmid><doi>10.1016/S0039-6060(97)90269-5</doi><tpages>10</tpages></addata></record> |
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| ispartof | Surgery, 1997-07, Vol.122 (1), p.91-100 |
| issn | 0039-6060 1532-7361 |
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| subjects | Adenoviridae - genetics Adult Base Sequence Biological and medical sciences Cell Count Cells, Cultured Electrophoresis, Polyacrylamide Gel Endothelium, Vascular - chemistry Endothelium, Vascular - cytology Endothelium, Vascular - metabolism Fibrin - analysis Gene Expression - physiology Gene Transfer Techniques - standards Genetic Vectors Humans Immunoblotting Medical sciences Miscellaneous Polytetrafluoroethylene Prostheses and Implants Pulsatile Flow Saphenous Vein - cytology Sodium Dodecyl Sulfate Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases Tissue Plasminogen Activator - analysis Tissue Plasminogen Activator - genetics Tissue Plasminogen Activator - metabolism |
| title | Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells |
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