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Ann Thorac Surg 1992;53:472-476
© 1992 The Society of Thoracic Surgeons


Articles

Flow cytometric analysis of organ preservation-induced endothelial cell membrane damage

William A. Killinger, MD, David B. Dorofi, BS, Ellis A. Tinsley, MD, Blair A. Keagy, MD, George Johnson, MD*

Department of Surgery, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, North Carolina USA

Accepted for publication August 30, 1991.

* Address reprint requests to Dr Johnson, UNC General Surgery, 210 Burnett-Womack, Campus Box 7210. Chapel Hill, NC 27599-7210 USA.

Organ preservation for transplantation is associated with endothelial cell damage. This vascular injury results in increased capillary permeability, graft edema, and early graft dysfunction. This damage may be the limiting factor in preservation of these organs. This study uses flow cytometric assessment of membrane integrity to examine the effects of various organ preservation solutions on human umbilical vein endothelial cell cultures. Confluent plates of human umbilical vein endothelial cells were incubated at 4 °C for 24, 48, and 72 hours in commonly used preservation solutions. After cold incubation, the cells were harvested and stained with propridium iodide and fluorescein diacetate. Cells were examined using a flow cytometer for membrane integrity and cytosolic activity. When examined after 24, 48, and 72 hours, cells stored at 4 °C in a 5% polyethylene glycol salt solution were significantly less damaged than those stored in any other solution (p < 0.05). After 48 and 72 hours at 4 °C, cells stored in ViaSpan were significantly more intact than cells stored in EuroCollins and 0.9% saline solution (p < 0.05). This study demonstrates that endothelial cell damage occurs during cold storage and that a polyethylene glycol-based solution showed superior cellular preservation.




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