CELL DETACHMENT DURING SINUSOIDAL REPERFUSION AFTER LIVER PRESERVATION - AN IN-VITRO MODEL

Background. Sinusoidal endothelial cells (SEC) are significantly more vulnerable to cold storage and reperfusion than hepatocytes. Swelling and disruption of the sinusoidal lining induce the microcirculatory di sturbances seen after reperfusion. In this article, the investigation of a method to assess the adhesion and morphology of SEC in vitro duri ng reperfusion after preservation is described. Methods. Time-lapse vi deo microscopy analysis was performed and cell detachment rates and ce ll lengths were determined, Preservation intervals between 6 and 24 hr and flow rates ranging from 3 L/min to 9 L/min (resulting in shear st resses between 5.1 and 15.3 dynes/cm(2) on the monolayer surface) duri ng reperfusion period were compared, SEC that were stored for 6 hr in University of Wisconsin solution and nonpreserved control cultures wer e compared. Results. Varying the preservation intervals from 6 hr to 2 4 hr during reperfusion at a flow rate of 3 L/min led to increased cel l erosion rates (6 hr, 35.5+/-15.2%; 12 hr, 38.0+/-7.6%; 18 hr, 54.3+/ -5.7%; 24 hr, 76.7+/-6.7%; non-preserved cells, 3.4+/-3.4%). Storage p eriods from 12 hr to 24 hr led to significantly higher cell detachment rates than occurred in nonpreserved cells. Conclusions. This method a llows the investigation of the adhesion capability and morphology of i ndividual cells in vitro. Indications of the kind of preservation/repe rfusion injury that occurs after treatment with several preservation s olutions and the resultant repair behavior can be obtained.