title: Cationic liposomal delivery of plasmid to endothelial cells measured by quantitative flow cytometry.
abstract: Cationic liposomes are potentially important gene transfer vehicles, capable of conjugating with anionic DNA by condensation. Flow cytometry was used to examine quantitatively the incorporation of DNA-liposome complex into murine capillary lung endothelial cells. The plasmid DNA, a pSV-beta-galactosidase vector, was covalently labeled with ethidium monoazide by photoactivation. The cationic liposome consisted of egg phosphatidylcholine (90%), cholesterol (5%), and stearylamine (5%). The number of plasmid molecules contained within each cell as a function of exposure time was estimated from fluorescence intensity. Fluorescently labeled plasmid is detectable after 10 min and increases with continued exposure, but at a decreasing rate, up to 2160 min. After 2160 min each cell, on average, contains approximately 10,000 plasmid molecules. Following transfection, a single cell unimodal population was detected by flow cytometry, suggesting that all cells participate in transfection equally. Furthermore, cell cycle analysis indicates that the entry of DNA-liposome complex is independent of cell cycle.
authors.affiliation: Department Chemical Engineering, Center for Intracellular Engineering, Vanderbilt University, Nashville, Tennessee 37235, USA.
authors.forename: W
authors.initials: W
authors.lastname: Tseng
authors.forename: N B
authors.initials: NB
authors.lastname: Purvis
authors.forename: F R
authors.initials: FR
authors.lastname: Haselton
authors.forename: T D
authors.initials: TD
authors.lastname: Giorgio