Endothelial cells are isolated from the saphenous vein segments. They are retrovirally transduced and expanded.
Endothelial cells (ECs) are isolated from the same saphenous vein segments of the patient. ECs are isolated using collagenase digestion.
4 x105 ECs are seeded on 60 mm ﬁbronectin-coated plates (4.5 lg/ml) 24 h prior to transduction. Transduction with recombinant retroviral vectors expressing the human Ang-1 is initiated by incubation with DEAE-dextran (1 mg/ml). The cells are then washed 3 times with PBS and exposed to the retroviral vectors in transduction medium for 4 h. The media containing the retroviruses is then replaced with growth medium. EC are cultured in the presence of G418 (0.5 mg/ml) for 2–4 days until at least 90% of the cells expressed the transduced genes.