title: DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes.
abstract: The effects of lipid compositions on their physicochemical properties and transfection efficiencies were investigated. Four liposome formulations with different 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) to dioleoylphosphatidylethanolamine (DOPE) weight ratios were investigated, that is, weight ratios 1:0 (T1P0), 3:1 (T3P1), 1:1 (T1P1), and 1:3 (T1P3). Mean sizes of liposomes were influenced by their lipid composition and the preparation concentration at the time of sonication. Zeta potentials of liposomes were inversely correlated with their liposome sizes. However, neither liposome sizes nor zeta potentials were correlated with transfection efficiency. The optimum composition of liposomes was cell-line dependent (T1P0 and T3P1 for Huh7 and AGS, T3P1 and T1P1 for COS7, and T1P1 and T1P3 for A549). The shape of lipoplexes was changed from lamellar to inverted hexagonal structure according to the increased ratio of DOPE, but there was no definite advantage of specific structure in transfection efficiency throughout all used cell lines. However, cellular internalization was consistently faster in T1P0, T3P1, T1P1 compared to T1P3 in all cell lines, suggesting the importance of endosomal escape. Our findings show that the transfection efficiency of DOTAP liposomes is mainly influenced by lipid composition and cell type, and not by size or zeta potential.
authors.affiliation: School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea; Department of Physiology, College of Medicine, Yeungnam University, Daegu 705-717, Republic of Korea.
authors.forename: Bieong-Kil
authors.initials: BK
authors.lastname: Kim
authors.affiliation: School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Guen-Bae
authors.initials: GB
authors.lastname: Hwang
authors.affiliation: School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Young-Bae
authors.initials: YB
authors.lastname: Seu
authors.affiliation: School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Jong-Soo
authors.initials: JS
authors.lastname: Choi
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, Republic of Korea.
authors.forename: Kyeong Sik
authors.initials: KS
authors.lastname: Jin
authors.affiliation: Department of Physiology, College of Medicine, Yeungnam University, Daegu 705-717, Republic of Korea.
authors.forename: Kyung-Oh
authors.initials: KO
authors.lastname: Doh
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D004058:Diffusion; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry*; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D008422:Materials Testing; D009374:Neoplasms, Experimental / Q000737:chemistry / Q000235:genetics*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
chemical_list: D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C094877:dioleoyl phosphatidylethanolamine; D004247:DNA; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Gene therapy; Liposomes; Transfection