Human microvessel endothelial cell isolation and vascular graft sodding in the operating room

Abstract

We have evaluated multiple factors inherent to an operating room-compatible endothelial cell procurement and sodding procedure. Microvessel endothelial cell isolations have been performed on fat tissue obtained from over 140 patients with a 100% success rate. Liposuction-derived fat was optimal with respect to cell yield, and isolation time. The devices and equipment used were acceptable to the operating room and the complete cell procurement procedure was successful even in the hands of personnel with minimal training. Fat digestion was achieved using crude clostridial collagenase, with an average cell yield of 1 x 10(6) microvessel endothelial cells/gm of fat. Evaluation of this procedure with canine fat using an operating room acceptable procedure resulted in a 100% procurement success rate requiring 1.5 hours (+/- .5 hrs) for completion of the fat isolation, and cell isolation procedure. Microvessel EC could subsequently be used in graft seeding or sodding techniques to establish endothelial cell monolayers on vascular grafts. Our results indicate that one person with minimal cell isolation background can reproducibly isolate large quantities of sterile autologous endothelial cells in the operating room for immediate use in endothelial cell seeding/sodding procedures.