Liposome-mediated high-efficiency transfection of human endothelial cells

Liposome-mediated transfection of endothelial cells provides a valuable exp erimental technique to study cellular gene expression and may also be adapt ed for gene therapy studies. However, the widely recognized disadvantage of liposome-mediated transfection is low efficiency. Therefore, studies were performed to optimize transfection techniques in human endothelial cells. T he majority of the experiments were performed with primary cultures of huma n umbilical vein endothelial cells (HUVEC). In addition, selected experimen ts were performed using human brain microvascular endothelial cells and hum an dermal microvascular endothelial cells. To study transfection rates, HUV EC were transfected with the pGL3 vector, containing the luciferase reporte r gene, complexed with several currently available liposomes, such as diffe rent Perfect Lipid (pFx) mixtures, DMRIE-C, or lipofectin. The optimal tran sfection rate was achieved in HUVEC transfected for 1.5 h with 5 mug/ml of DNA plasmid in the presence of 36 mug/ml of pFx-7, In addition, transfectio n with the VR-3301 vector encoding for human placental alkaline phosphatase revealed that, under the described conditions, transfection efficiency in HUVEC was approximately 32%, Transfections mediated by other liposomes were less efficient. The usefulness of the optimized transfection technique was confirmed in HUVEC transfected with NF-kappaB or AP-1-responsive construct s and stimulated with TNF or LPS. We conclude that among several currently available liposomes, pFx-7 appears to be the most suitable for transfection s of cultured human endothelial cells. Copyright (C) 2001 S. Karger AG, Bas el.