Transfection of Small Numbers of Human Endothelial Cells by Electroporation and Synthetic Amphiphiles

Abstract 

Objectives:this study compared the efficiency of electroporation and synthetic amphiphiles (SAINT-2^pp/DOPE) in transfecting small numbers of human endothelial cells.Methods and results:optimal transfection conditions were tested and appeared to be 400 V and 960 μF for electroporation and a 10:1 ratio for concentrations of SAINT-2^pp/DOPE: plasmid. Using these conditions, cell concentrations were lowered step-wise and we were able to transfect as few as one thousand cells with both methods. For detection of transfection of a small number of cells a sensitive assay was needed (Luciferase). A plasmid containing the neomycin resistance gene was used to determine the transfection rate expressed in colony forming units by counting colonies after selection. At low plasmid concentrations this transfection rate was within the same range for both electroporation and SAINT-2^pp/DOPE transfection. Fluorescentin situhybridisation of metaphase chromosomes of transfected endothelial cells using the plasmid as a probe showed that stable integration was possible with both methods.Conclusions:electroporation and a synthetic amphiphile, SAINT-2^pp, provide the possibility of transfecting small numbers of cells resulting in stable integration of low plasmid concentrations. The availability of this technology is important in order to obtain functional endothelial cell lines from various human blood vessels for research purposes.

Keywords: Electroporation, Endothelial cells, Lipofection, Synthetic amphiphile, Thrombosis, Transfection.

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• f1 Please address all correspondence to: M. H. J. Ruiters, Centre for Biomedical Technology, University of Groningen, PO Box 145, 9700 AC Groningen, The Netherlands.