1. Transfection efficiency and cytotoxicity of nonviral gene transfer reagents in human smooth muscle and endothelial cells.
- Author
-
Kiefer K, Clement J, Garidel P, and Peschka-Süss R
- Subjects
- Animals, Aorta pathology, Cell Line, Chemistry, Pharmaceutical methods, Chemistry, Pharmaceutical standards, Cholesterol chemistry, Cholesterol pharmacology, DNA administration & dosage, Drug Combinations, Drug Evaluation, Preclinical methods, Endothelial Cells chemistry, Endothelium, Vascular metabolism, Genetic Therapy methods, Germany, Humans, Indicators and Reagents classification, Indicators and Reagents pharmacology, Luminescent Proteins chemistry, Luminescent Proteins pharmacology, Muscle, Smooth, Vascular metabolism, Phosphatidylethanolamines chemistry, Phosphatidylethanolamines pharmacology, Plasmids genetics, Quality Control, Rats, Transfection methods, United States, United States Food and Drug Administration legislation & jurisprudence, Cell Survival, Cholesterol analogs & derivatives, Endothelium, Vascular cytology, Gene Transfer Techniques, Indicators and Reagents chemistry, Muscle, Smooth, Vascular cytology, Transfection standards
- Abstract
Purpose: Evaluation of a nonviral transfection reagent with respect to efficient gene transfer into primary human vascular cells., Methods: Complexes consisting of seven commercially available transfection reagents (DAC-30, DC-30, Lipofectin, LipofectAMINE PLUS, Effectene, FuGene 6 and Superfect) and EGFP encoding plasmid DNA were studied. The in vitro transfection efficiency and cytotoxicity in human aorta smooth muscle cells (HASMCs) and endothelial cells (HAECs) and rat smooth muscle cells (A-10 SMCs) were assayed in the presence of serum using flow cytometric analysis and ATP-quantitation assay, respectively., Results: Human primary cells were transfected less efficiently compared to the rat smooth muscle cell line. Transfection efficiency depended on the type of reagent, the reagent/DNA ratio, and, most importantly, on the cell type used. Determination of cytotoxicity showed that the effects of transfection on cell viability did not significantly differ from one another depending on the cell type. The exception to this was Superfect, which obviously reduced cell viability in all cell types., Conclusions: Our experiments showed that DAC-30 is the preferred transfection reagent for HASMCs and HAECs, exhibiting an improved efficiency combined with an acceptable cytotoxicity. Therefore, it might offer a therapeutic option for the treatment of cardiovascular disease and prove suitable for further drug development.
- Published
- 2004
- Full Text
- View/download PDF