MultiGeneAngio, transduced vein-derived endothelial cells (Multi Gene Vascular Systems (MGVS))

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.