The Study On The Tissue Engineering Homograft Valve

Part 1. Cultivation Of The Adult Human Endothelium From The Great Saphenous VeinObjective: To investigate the suitable method to cultivate the adult human endothelium from the great saphenous vein .Method: Endothelial cells(EC) from small discarded segments of human great saphenous vein of patients undergoing coronary artery bypass surgery. EC were digested with 2% collagenase II in medium 199, seeded into culture flask with high density about ( 1.5 × 10⁴ /cm² ), after reaching confluence ,the EC were detached with a 0.25% trypsin-ethylenediamine tetraacetic acid (EDTA, 1 mmol·L-1) solution ,and seeded into culture flask with low density about ( 0.8 × 10⁴ /cm²) ,subcultivated as described for primary cultures (split ratio 1:3).Results: Primary endothelial cells were confluenced about (12 ±2.3 ) d and 7.0×10⁴ /cm² EC or more could be achieved, the growth of passage cells were more exuberant and confluenced about (7.0 ±1.1) d. the EC retained spindle-shaped or cobblestone appearance,Weibel-Palade(WP) could be found by the transmission electron microscopy, and CD₃₁ antigen were positive in the EC by the immunofluorescent histochemistry.Conclusions: We suggest that this provides a practical useful method for study of cultured endothelial cells and tissue engineering.Part 2. The Study On The Decellular Human Homograft Aortic Valves Conduits.