title: Differential behaviour of lipid based and polycation based gene transfer systems in transfecting primary human fibroblasts: a potential role of polylysine in nuclear transport.
abstract: DNA delivery systems for gene therapy applications have to be able to trigger the uptake of plasmid DNA into the nucleus. We have tested two types of non-viral vector systems, lipofection (cationic lipid-based, using Lipofectamine) and polyfection (cationic polymer-based, using glycerol enhanced transferrinfection), for their ability to transfect confluent, contact inhibited primary human fibroblasts. While both systems worked well with growing fibroblasts, polyfection was superior with confluent cells. A slight reduction in cell associated plasmid DNA was observed with resting cells, but it was similar for both types of complexes. Lipofectamine showed a prevalence for transfecting cycling cells as judged by costaining transfected cells with cell cycle markers. No such bias was observed when glycerol enhanced transferrinfection was used. Microinjection of plasmid DNA/polylysine complexes into the cytoplasm of fibroblasts resulted in a higher percentage of expressing cells than injection of plasmid DNA, offering an explanation for the higher transfection levels obtained with transferrinfection in non-growing cells.
authors.affiliation: Institute of Biochemistry, University of Vienna, Dr. Bohrgasse 9/3, A-1030, Vienna, Austria. wolfgang.zauner@cobrat.com
authors.forename: W
authors.initials: W
authors.lastname: Zauner
authors.forename: S
authors.initials: S
authors.lastname: Brunner
authors.forename: M
authors.initials: M
authors.lastname: Buschle
authors.forename: M
authors.initials: M
authors.lastname: Ogris
authors.forename: E
authors.initials: E
authors.lastname: Wagner
mesh_terms: D001692:Biological Transport; D002411:Cation Exchange Resins; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D015316:Genetic Therapy; D006801:Humans; D008055:Lipids; D008156:Luciferases; D008845:Microinjections; D010957:Plasmids; D011107:Polylysine / Q000031:analogs & derivatives* / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014168:Transferrin / Q000031:analogs & derivatives*
chemical_list: D002411:Cation Exchange Resins; D008055:Lipids; C086724:Lipofectamine; D014168:Transferrin; C072481:transferrin-polylysine conjugate; D011107:Polylysine; D008156:Luciferases