title: Gene transfer into specific vascular cells.
abstract: Gene transfer therapy in vascular surgery is on the horizon and will include the insertion of genes for anti-clotting proteins into the endothelial lining of vascular grafts and for genes controlling the proliferation of smooth muscle cells after endovascular intervention. Here we address the possibility of targeting genes to specific vascular cells using non-infectious methods, DEAE-dextran or lipofectin complexes of reporter genes, to aid transfection of endothelial cells, smooth muscle cells and fibroblasts cultured from human umbilical veins or arteries. For these studies we used the firefly luciferase gene under control of several different promoters including those for the Rous sarcoma virus (RSV) and for tissue plasminogen activator type 1 (PAI-1). DEAE-dextran mediated transfections resulted in low level, transient (2-5 days) expression of RSV-luciferase in all three cell types. Lipofectin mediated transfections resulted in a four-to-five-fold higher expression of RSV-luciferase in endothelial and smooth muscle cells, expression remaining fairly stable for up to 14 days. One particular PAI-1 promoter construct of 800 bp was only half as effective as the RSV promoter in the expression of luciferase from smooth muscle cells, 82 +/- 9 and 35 +/- 11 ng mg-1 respectively (p less than 0.02). In contrast these two promoters resulted in very similar expression of luciferase in endothelial cells, 64 +/- 8 and 67 +/- 10 ng mg-1 respectively. These experiments demonstrate the possibilities of augmenting cultured vascular cells with foreign genes using lipofectin, a cationic lipid, for insertions into endothelial and smooth muscle cells.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Central Laboratory of The Netherlands Red Cross, Amsterdam.
authors.forename: J T
authors.initials: JT
authors.lastname: Powell
authors.forename: A J
authors.initials: AJ
authors.lastname: van Zonneveld
authors.forename: J A
authors.initials: JA
authors.lastname: van Mourik
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D001517:Coleoptera / Q000235:genetics*; D003637:DEAE-Dextran; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D010714:Phosphatidylethanolamines; D014162:Transfection
chemical_list: D010714:Phosphatidylethanolamines; C020888:1,2-dielaidoylphosphatidylethanolamine; D003637:DEAE-Dextran; D008156:Luciferases