Magnetofection Potentiates Gene Delivery to Cultured Endothelial Cells
| Autor: | Ulrich Pohl, Hae-Young Sohn, Torsten Gloe, Florian Krötz, Christian Plank |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Endothelial cells
Magnetofection Plasmid vectors Swine Physiology viruses Gene delivery Biology Transfection Green fluorescent protein Magnetics Animals Humans Polyethyleneimine Luciferase Luciferases Cells Cultured L-Lactate Dehydrogenase Genetic transfer Endothelial Cells Molecular biology ddc Endothelial stem cell Cell culture Gene Targeting embryonic structures Cardiology and Cardiovascular Medicine |
| Zdroj: | Journal of Vascular Research. 40:425-434 |
| ISSN: | 1423-0135 1018-1172 |
| DOI: | 10.1159/000073901 |
| Popis: | 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 a β-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. |
| Databáze: | OpenAIRE |
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