Abstract
Canine vascular endothelial cells (CECs) were obtained by collagenase perfusion from the external jugular vein and cultured serially for the purpose of applying to the prostheses covered with endothelial cells (cultured graft). We cultured the CECs on ePTFE sheets and determined their growth requirements. On native culture dishes, CECs grew rapidly in nutrient media supplemented with 10% fetal bovine serum, EGF (10ng/ml), insulin (1ng/ml), transferrin (10μg/ml), acidic fibroblast growth factor (aFGF, 20ng/ml) and heparin (25μg/ml). The aFGF was essential for CECs' growth and type 1 collagen coating of culture dishes enhanced their proliferation. When culture on ePTFE sheets, CECs were neither able to adhere nor grow on ePTFE alone (the rate of attachement 4.5%). However, they adhered and grew well on collagen coated sheets. The optimal concentration of collagen was 25μg/ml, the rate of attachement was 30-50%, and the doubling time was 30-35 hours. When inoculated at 180cells/mm2, CECs grew to confluence after around 5 day-incubation in culture, and scanning electron microscopy and light microscopy showed CECs monolayer on the ePTFE sheets. Thus, ePTFE sheets coated with collagen might be useful as a material to make a vascular graft covered with endothelial cells. We showed here the basic conditions for the making of cultured tube graft.
