title: Physico-chemical characterization and transfection efficacy of cationic liposomes containing the pEGFP plasmid.
abstract: Cationic liposomes-DNA complexes (lipoplexes) are largely used in gene delivery. Deciphering specific chemical and physical properties of lipoplexes is a necessary step to unravel the mechanisms underlying transfection and to improve transfection efficacy in each experimental model. In the present paper we investigated the physico-chemical features of lipoplexes containing a plasmid encoding for the GFP protein, in order to correlate these results with transfection efficacy. Cationic unilamellar vesicles (mean diameter 100 nm) were prepared, from the cationic DC-Chol lipid and the zwitterionic phospholipid DOPE. The two components of the liposome bilayer were used at molar ratio close to unity. ESR spectra were recorded and zeta potential zeta was measured on liposomes complexed with the plasmid. One of the main points of interest in this paper resided in the fact that both kinds of measurements were carried out in the same conditions (i.e. lipid concentration, medium composition, and pH) employed for cell transfection experiments. Transfection was performed on CHO cells; the percentage of fluorescent cells was evaluated and compared with the above physico-chemical features. It emerged that the composition and pH of the medium, the lipoplex/cell ratio, as well as the amount of lipoplex added to the cell culture were critical parameters for transfection efficacy. Finally, lipoplex surface charge played a fundamental role to achieve a high transfection level.
authors.affiliation: Department of Chemistry & CSGI, University of Firenze, 50019 Sesto F.no, Firenze, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Salvati
authors.forename: Laura
authors.initials: L
authors.lastname: Ciani
authors.forename: Sandra
authors.initials: S
authors.lastname: Ristori
authors.forename: Giacomo
authors.initials: G
authors.lastname: Martini
authors.forename: Alessio
authors.initials: A
authors.lastname: Masi
authors.forename: Annarosa
authors.initials: A
authors.lastname: Arcangeli
mesh_terms: D000818:Animals; D016466:CHO Cells; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D010957:Plasmids; D014162:Transfection
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D008081:Liposomes; D010714:Phosphatidylethanolamines; C485184:enhanced green fluorescent protein; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; D049452:Green Fluorescent Proteins; D004247:DNA; D002784:Cholesterol