Transfection efficiency and cytotoxicity of nonviral gene transfer reagents in human smooth muscle and endothelial cells.
OBJECTIVE
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.
Kiefer
Karin
K
Department of Pharmaceutical Technology, Albert-Ludwigs University, D-79104 Freiburg, Germany.
Clement
Jule
J
Garidel
Patrick
P
Peschka-Süss
Regine
R
D000818:Animals; D001011:Aorta / Q000473:pathology; D002460:Cell Line; D002470:Cell Survival; D002626:Chemistry, Pharmaceutical / Q000379:methods / Q000592:standards; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D004247:DNA / Q000008:administration & dosage; D004338:Drug Combinations; D004353:Drug Evaluation, Preclinical / Q000379:methods; D042783:Endothelial Cells / Q000737:chemistry; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005858:Germany; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry* / Q000145:classification / Q000494:pharmacology; D008164:Luminescent Proteins / Q000737:chemistry / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011786:Quality Control; D051381:Rats; D014162:Transfection / Q000379:methods / Q000592:standards*; D014481:United States; D014486:United States Food and Drug Administration / Q000331:legislation & jurisprudence
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D004338:Drug Combinations; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D010714:Phosphatidylethanolamines; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; D004247:DNA; D002784:Cholesterol