In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system
To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells. Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylchol...
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| Veröffentlicht in: | Pharmaceutical research 2003-05, Vol.20 (5), p.738-744 |
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| creator | GUANGLIANG PAN SHAWER, Mohannad ØIE, Svein LU, D. Robert |
| description | To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells.
Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system.
The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system.
A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system. |
| doi_str_mv | 10.1023/A:1023477317668 |
| format | Article |
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Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system.
The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system.
A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system.</description><identifier>ISSN: 0724-8741</identifier><identifier>EISSN: 1573-904X</identifier><identifier>DOI: 10.1023/A:1023477317668</identifier><identifier>PMID: 12751628</identifier><identifier>CODEN: PHREEB</identifier><language>eng</language><publisher>New York, NY: Springer</publisher><subject>Biological and medical sciences ; Cell Line, Tumor - drug effects ; Cholesterol ; Cytotoxicity ; Drug Delivery Systems - methods ; Efficiency ; General pharmacology ; Genetic engineering ; Genetic Therapy - methods ; Glioma - drug therapy ; Glioma - genetics ; Humans ; Lipids ; Lipoproteins ; Lipoproteins - administration & dosage ; Lipoproteins - genetics ; Lipoproteins - toxicity ; Medical sciences ; Nanoemulsions ; Pharmaceutical technology. Pharmaceutical industry ; Pharmacology. Drug treatments ; Proteins ; Transfection - methods ; Vectors (Biology)</subject><ispartof>Pharmaceutical research, 2003-05, Vol.20 (5), p.738-744</ispartof><rights>2003 INIST-CNRS</rights><rights>Copyright Kluwer Academic Publishers May 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c310t-4755d9a6afc7c60c5067d975dd3d7bed34f40c35daeaf724a1002aed2f69cbf43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,778,782,27907,27908</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14771843$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12751628$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>GUANGLIANG PAN</creatorcontrib><creatorcontrib>SHAWER, Mohannad</creatorcontrib><creatorcontrib>ØIE, Svein</creatorcontrib><creatorcontrib>LU, D. Robert</creatorcontrib><title>In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system</title><title>Pharmaceutical research</title><addtitle>Pharm Res</addtitle><description>To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells.
Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system.
The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system.
A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system.</description><subject>Biological and medical sciences</subject><subject>Cell Line, Tumor - drug effects</subject><subject>Cholesterol</subject><subject>Cytotoxicity</subject><subject>Drug Delivery Systems - methods</subject><subject>Efficiency</subject><subject>General pharmacology</subject><subject>Genetic engineering</subject><subject>Genetic Therapy - methods</subject><subject>Glioma - drug therapy</subject><subject>Glioma - genetics</subject><subject>Humans</subject><subject>Lipids</subject><subject>Lipoproteins</subject><subject>Lipoproteins - administration & dosage</subject><subject>Lipoproteins - genetics</subject><subject>Lipoproteins - toxicity</subject><subject>Medical sciences</subject><subject>Nanoemulsions</subject><subject>Pharmaceutical technology. Pharmaceutical industry</subject><subject>Pharmacology. Drug treatments</subject><subject>Proteins</subject><subject>Transfection - methods</subject><subject>Vectors (Biology)</subject><issn>0724-8741</issn><issn>1573-904X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNpFkUtLAzEUhYMotlbX7iQILkfzmsmMOyk-CgU3Cu7KbR41JZPUSaa0_94RK67O5uPce85B6JKSW0oYv3u4_xEhJaeyquojNKal5EVDxMcxGhPJRFFLQUfoLKU1IaSmjThFI8pkSStWj9FuFvDW5S7ilQkG5w5CskZlFwN2AX_2LQS88i62gJXxPuE-ubDCgEPcGo8haOxNSljtc8xx5xSGLjvrlAOPvdvETRezGay08W5ruj1O-5RNe45OLPhkLg46Qe9Pj2_Tl2L--jybPswLxSnJhZBlqRuowCqpKqJKUkndyFJrruXSaC6sIIqXGgzYIS5QQhgYzWzVqKUVfIKuf32HP756k_JiHfsuDCcXjBEqJWVkgK4OUL9sjV5sOtdCt1_89TQANwcAkgJvh5qUS__csACtBeffkSZ5nw</recordid><startdate>20030501</startdate><enddate>20030501</enddate><creator>GUANGLIANG PAN</creator><creator>SHAWER, Mohannad</creator><creator>ØIE, Svein</creator><creator>LU, D. Robert</creator><general>Springer</general><general>Springer Nature B.V</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>3V.</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>20030501</creationdate><title>In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system</title><author>GUANGLIANG PAN ; SHAWER, Mohannad ; ØIE, Svein ; LU, D. Robert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c310t-4755d9a6afc7c60c5067d975dd3d7bed34f40c35daeaf724a1002aed2f69cbf43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Biological and medical sciences</topic><topic>Cell Line, Tumor - drug effects</topic><topic>Cholesterol</topic><topic>Cytotoxicity</topic><topic>Drug Delivery Systems - methods</topic><topic>Efficiency</topic><topic>General pharmacology</topic><topic>Genetic engineering</topic><topic>Genetic Therapy - methods</topic><topic>Glioma - drug therapy</topic><topic>Glioma - genetics</topic><topic>Humans</topic><topic>Lipids</topic><topic>Lipoproteins</topic><topic>Lipoproteins - administration & dosage</topic><topic>Lipoproteins - genetics</topic><topic>Lipoproteins - toxicity</topic><topic>Medical sciences</topic><topic>Nanoemulsions</topic><topic>Pharmaceutical technology. Pharmaceutical industry</topic><topic>Pharmacology. Drug treatments</topic><topic>Proteins</topic><topic>Transfection - methods</topic><topic>Vectors (Biology)</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GUANGLIANG PAN</creatorcontrib><creatorcontrib>SHAWER, Mohannad</creatorcontrib><creatorcontrib>ØIE, Svein</creatorcontrib><creatorcontrib>LU, D. Robert</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>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><jtitle>Pharmaceutical research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>GUANGLIANG PAN</au><au>SHAWER, Mohannad</au><au>ØIE, Svein</au><au>LU, D. Robert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system</atitle><jtitle>Pharmaceutical research</jtitle><addtitle>Pharm Res</addtitle><date>2003-05-01</date><risdate>2003</risdate><volume>20</volume><issue>5</issue><spage>738</spage><epage>744</epage><pages>738-744</pages><issn>0724-8741</issn><eissn>1573-904X</eissn><coden>PHREEB</coden><abstract>To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells.
Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system.
The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system.
A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system.</abstract><cop>New York, NY</cop><pub>Springer</pub><pmid>12751628</pmid><doi>10.1023/A:1023477317668</doi><tpages>7</tpages></addata></record> |
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| ispartof | Pharmaceutical research, 2003-05, Vol.20 (5), p.738-744 |
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| subjects | Biological and medical sciences Cell Line, Tumor - drug effects Cholesterol Cytotoxicity Drug Delivery Systems - methods Efficiency General pharmacology Genetic engineering Genetic Therapy - methods Glioma - drug therapy Glioma - genetics Humans Lipids Lipoproteins Lipoproteins - administration & dosage Lipoproteins - genetics Lipoproteins - toxicity Medical sciences Nanoemulsions Pharmaceutical technology. Pharmaceutical industry Pharmacology. Drug treatments Proteins Transfection - methods Vectors (Biology) |
| title | In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system |
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