Endothelial cell harvest for seeding vascular prostheses: the influence of technique on cell function, viability, and number.

Direct seeding of endothelial cells onto synthetic vascular prostheses has become the subject of increasing surgical research during the last 5 to 7 years. The currently employed cell harvest techniques are inefficient, resulting in cell counts far below the number of cells calculated to be present on the original donor vein. We have compared two methods of enzymatic endothelial cell harvest: cannulation with flushing and eversion over a stainless steel rod. Harvested cells were plated onto tissue culture plastic and counted after 24 hours of incubation. The methods ensured that only those cells viable and functional enough to adhere to the plastic were being considered. Cells were identified as endothelial by immunohistochemical techniques applying antisera to factor VIII-related antigen. Segments of normal vein and of veins treated by each technique were viewed with a scanning electron microscope. Cannulation was the superior method, providing greater numbers of viable, functional cells. The eversion technique was unreliable and probably injurious to endothelial cells.