Lipoplex formation using liposomes prepared by ethanol injection.
Cationic liposomes composed of 3beta-[N-(N'N'-dimethylaminoethane)carbamoyl] cholesterol (DC-Chol) and dioleoylphosphatidylethanolamine (DOPE) (DC-Chol/DOPE liposome, molar ratio, 1:1 or 3:2) prepared by the dry-film method have been often used as non-viral gene delivery vectors. We have shown that a more efficient transfection in medium with serum was achieved using DC-Chol/DOPE liposomes (molar ratio, 1:2) than those (3:2), and preparation method by a modified ethanol injection than the dry-film. The most efficient DC-Chol/DOPE liposome for gene transfer was molar ratio (1:2) and prepared by a modified ethanol injection method. The enhanced transfection is related to an increase in the release of DNA in the cytoplasm by the large lipoplex during incubation in opti-MEM I reduced-serum medium (optiMEM), not to an increased cellular association with the lipoplex. Cationic liposomes rich in DOPE prepared by a modified ethanol injection method will help to improve the efficacy of liposome vector systems for gene delivery.
Maitani
Yoshie
Y
Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
D002412:Cations / Q000737:chemistry; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage*; D000431:Ethanol; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D008156:Luciferases / Q000235:genetics; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D014162:Transfection
D002412:Cations; D008081:Liposomes; D010714:Phosphatidylethanolamines; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C094877:dioleoyl phosphatidylethanolamine; D000431:Ethanol; D004247:DNA; D002784:Cholesterol; D008156:Luciferases