High‐efficiency transfection of human endothelial cells mediated by cationic lipids

Cellular transfections provide powerful experimental tools to study gene regulation. However, endothelial cells are difficult to transfect. Therefore, the present studies were performed to optimize transfection techniques in human endothelial cells. To study transfection rates, endothelial cells were transfected with the pGL3 vector, containing the luciferase reporter gene, complexed with several currently available liposomes, such as different Perfect Lipid (pFx) mixtures, DMRIE‐C, or lipofectin. The optimal transfection rate was achieved in cells transfected for 1.5 h with 5 mg/mL of DNA plasmid in the presence of 36 mg/mL of pFx‐7. Transfections mediated by other liposomes were less efficient. Usefulness of the optimized transfection technique was confirmed in cells transfected with NF‐kB or AP‐1‐responsive constructs. We conclude that among several currently available liposomes, pFx‐7 appears to be the most suitable for transfections of cultured human endothelial cells. Acknowledgements: Supported by NIH; NS39254, MH63022, and AA013843.