Seeding And Expression Of Gene-Modified Endothelial Cells On Small Diameter Vascular Graft In Vitro

There is still no successful clinical use of small diameter vascular graft so far after the first report of ECs seeded on vascular prostheses in canine by Herring in 1978. The main limitaiton is early thrombosis after implanting caused by endothelialization only in part. These are relating to not only the biological characteristics of ECs, but also the method of ECs seeding and thrombolytic therapy. In this research, we studied the ECs specially for seeding in cellular level, created 'compressed-rotated seeding' method and finally transferred the human pro-UK cDNA into the ECs seeded on small diameter vascular graft throuth adenovirus vector. The gene expression was detected by Western blot in vitro.Our purpose of the first part of this study was to study what kind of ECs are better for seeding on vascular grafts. First, we digested cells from the intima of human umbilical vein vessel. These cells were confirmed to be endothelial cells by specific factor â…§ related antigen stained with immunofluorescence. And then the studies were taken as follows:1. Comparing collagenase with trypsin: 6 umbilical cords were divided into 2 groups. Collagenase group (3 cords) were digested using 0.1% collagenase. Trypsin group were digested using 0.25% trypsin. The results revealed that the days of collagenase group from seeding to...