Magnetofection potentiates gene delivery to cultured endothelial cells

被引:99
作者
Krötz, F
Sohn, HY
Gloe, T
Plank, C
Pohl, U
机构
[1] Univ Munich, Inst Physiol, DE-80336 Munich, Germany
[2] Univ Munich, Div Cardiol, Med Poliklin, Innenstadt, DE-80336 Munich, Germany
[3] Tech Univ Munich, Dept Expt Oncol, D-8000 Munich, Germany
关键词
endothelial cells; magnetofection; plasmid vectors;
D O I
10.1159/000073901
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Modification of cellular functions by overexpression of genes is increasingly practised for research of signalling pathways, but restricted by limitations of low efficiency. We investigated whether the novel technique of magnetofection (MF) could enhance gene transfer to cultured primary endothelial cells. MF of human umbilical vein endothelial cells (HUVEC) increased transfection efficiency of a luciferase reporter gene up to 360-fold compared to various conventional transfection systems. In contrast, there was only an up to 1.6-fold increase in toxicity caused by MF suggesting that the advantages of MF outbalanced the increase in toxicity. MF efficiently increased transfection efficiency using several commercially available cationic lipid transfection reagents and polyethyleneimine (PEI). Using PEI, even confluent HUVEC could be efficiently transfected to express luciferase activity. Using a green fluorescent protein vector maximum percentages of transfected cells amounted up to 38.7% while PEI without MF resulted in only 1.3% transfected cells. Likewise, in porcine aortic endothelial cells MF increased expression of a luciferase or beta-galactosidase reporter, reaching an efficiency of 37.5% of cells. MF is an effective tool for pDNA transfection of endothelial cells allowing high efficiencies. It may be of great use for investigating protein function in cell culture experiments.
引用
收藏
页码:425 / 434
页数:10
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