Transfection of small numbers of human endothelial cells by electroporation and synthetic amphiphiles

Objectives: this study compared rite efficiency of electroporation and synt hetic amphiphiles (SAINT-2(pp)/DOPE) in transfecting small numbers of human endothelial cells. Methods and results: optimal transfection conditions were tested and appear ed to be 400 V and 960 mu F for electroporation and a 10:1 ratio for concen trations of SAINT-2(pp)/DOPE: plasmid. Using these conditions, cell concent rations were lowered step-wise and we were able to transfect as Jew 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 contai ning the neomycin resistance gene was used to determine the transfection ra te expressed in colony forming units by counting colonies after selection. At low plasmid concentrations this transfection rate was within the same ra nge for both electroporation and SAINT-2(pp)/DOPE transfection. Fluorescent in situ hybridisation of metaphase chromosomes of transfected endothelial cells using the plasmid as a probe showed that stable integration was possi ble with both methods. Conclusions: electroporation and a synthetic amphiphile, SAINT-2(pp), provi de the possibility of transfecting small numbers of cells resulting in stab le integration of low plasmid concentrations. The availability of this tech nology is important in ol der to obtain functional endothelial cell lines f rom various human blood vessels for research purposes.