Transfection Protocol for Human Umbilical Vein Endothelial Cells ( HUVEC) and Human Microvascular Endothelial Cells ( HMVEC ) in Targefect Handbook of Transfection Protocols

Abstract

Efficient gene transfer into primary endothelial cells is an important prerequisite for studying regulation of gene expression in vascular tissue. Here we present a simple set of protocols using the Targefect-HUVEC TM transfection kit to transfect HUVECs, HMVECs, BAECs and different types of primary endothelial cells. The kit consists of Targefect F-2 , a non-lipid cationic transfection reagent with low toxicity along with two enhancer formulations- Virofect TM and a Peptide Enhancer. Virofect enhances gene transfer by using adenoviral receptors on the cell surface to enhance intracellular delivery of transfection complexes. Once the transfection complex is internalized, Virofect helps the transfection complex escape degradation in the lysosome and enhances the both duration and efficiency of transgene expression. The Peptide Enhancer increases efficiency of transgene expression by escorting genes to the nucleus. Using the protocols mentioned here researchers have been able to achieve 70-80% transfection efficiencies in HUVECs and efficiencies ranging form 70-80% in human and bovine lung microvascular cells (see attached figures showing data from other lab groups and a list of product citations http://www.targetingsystems.net/pdf/Targefect_HUVEC.pdf )