ASSESSMENT OF ENDOTHELIAL PRESERVATION IN HUMAN CELL-CULTURES

Background. Impairment of microcirculation due to endothelial cell dam age must be considered a limiting factor in organ preservation. The pr esent study aims at a quantitative assessment of preservation-induced injury in cultured human endothelial cells. Methods. Monolayer culture s of human umbilical vein endothelial cells were exposed to cold (4 de grees C) hypoxic storage in University of Wisconsin solution, histidin e-tryptophane-ketoglutarate solution, Euro-Collins solution, and salin e solution. Cellular integrity was evaluated by viable cell count, ult rastructural analysis, and prostacyclin release after 24, 48, and 72 h ours of storage and subsequent 6 hours of reincubation in culture medi um at 37 degrees C. Expression of intercellular adhesion molecule-1 wa s investigated after 6, 12 and 24 hours of cold preservation and after 6 hours of rewarming. Results. Cellular viability was best maintained with University of Wisconsin and histidine-tryptophane-ketoglutarate solutions with no significant reduction of cell count up to 72 hours; Euro-Collins solution and saline solution caused a significant decline in cell numbers after 24 hours (p < 0.05). Morphology was best preser ved by University of Wisconsin solution. Prostacyclin values were elev ated after 24 hours in Euro-Collins solution and saline solution, afte r 48 hours in histidine-tryptophane-ketoglutarate, Euro-Collins, and s aline solutions, and after 72 hours in Euro-Collins solution (p < 0.05 , compared with University of Wisconsin solution). ICAM expression was weak after cold storage (24 hours) in University of Wisconsin solutio n, moderate after incubation in histidine-tryptophane-ketoglutarate an d Euro-Collins solutions and intensive after storage in saline solutio n. In contrast, rewarming caused intensive expression of intercellular adhesion molecule-1 in all experimental groups as compared with contr ols, which showed baseline expression at any time. Conclusions. From o ur results we conclude that in this model cellular integrity is best p rotected by University of Wisconsin solution, increased prostacyclin r elease is consistent with morphologic alterations and intercellular ad hesion molecule-1 expression is clearly up-regulated in endothelial ce lls under reperfusion conditions after cold hypoxic storage.