Flow cytometric analysis of organ preservation-induced endothelial cell membrane damage.
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 degrees 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 degrees C in a 5% polyethylene glycol salt solution were significantly less damaged than those stored in any other solution (p less than 0.05). After 48 and 72 hours at 4 degrees C, cells stored in ViaSpan were significantly more intact than cells stored in EuroCollins and 0.9% saline solution (p less than 0.05). This study demonstrates that endothelial cell damage occurs during cold storage and that a polyethylene glycol-based solution showed superior cellular preservation.
Killinger
W A
WA
Department of Surgery, University of North Carolina, Chapel Hill School of Medicine.
Dorofi
D B
DB
Tinsley
E A
EA
Keagy
B A
BA
Johnson
G
G
D000241:Adenosine; D000493:Allopurinol; D002470:Cell Survival; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000473:pathology*; D005434:Flow Cytometry; D005978:Glutathione; D006801:Humans; D006982:Hypertonic Solutions / Q000633:toxicity; D007328:Insulin; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D011092:Polyethylene Glycols / Q000633:toxicity; D011887:Raffinose; D012965:Sodium Chloride / Q000633:toxicity; D012996:Solutions / Q000633:toxicity; D013696:Temperature; D014471:Umbilical Veins / Q000473:pathology
C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; D012996:Solutions; C055206:University of Wisconsin-lactobionate solution; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose