High-efficiency transformation of human endothelial cells by Apo E-mediated transfection with plasmid DNA.
Endothelial cells, because of their proximity to the blood stream, provide an attractive system for gene transfer and delivery of gene products that control foci of vascular disease processes. We describe a simple, new methodology to achieve highly efficient transformation of cultured human endothelial cells derived from umbilical veins (HUVEC). A plasmid pCH110 containing coding region for beta-galactosidase driven by SV 40 early promoter region was employed to transfect HUVEC. The developed protocol exploits the role of apolipoprotein E (Apo E) in the metabolism of Apo E-containing lipoproteins and its high affinity binding to LDL receptors. DNA transfection of cultured HUVEC was carried out using standard transfection methods including calcium phosphate precipitation, polybrene mediated transfection, and lipofection. The new methodology of transfecting HUVEC employed Apo E adsorbed lipofection reagent-DNA complex, and was found to be the most efficient procedure to transform HUVEC in comparison to the standard methods used in this study.
Sipehia
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R
Artificial Cells and Organs Research Centre, Faculty of Medicine, McGill University, Montreal, Quebec, Canada.
Martucci
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G
D001057:Apolipoproteins E / Q000378:metabolism*; D002478:Cells, Cultured; D004274:DNA, Recombinant; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D010957:Plasmids; D011485:Protein Binding; D014162:Transfection; D014170:Transformation, Genetic
D001057:Apolipoproteins E; D004274:DNA, Recombinant