[Preservation of hepatocytes in UW and HTK solutions with added fluorocarbons]. / Preservación de hepatocitos en soluciones de UW y HTK con adición de fluorocarbonos.

ABSTRACT

BACKGROUND: One of the limitations for hepatocyte transplantation is the short survival of isolated hepatocytes. OBJECTIVE: We evaluated the utility of fluorocarbons (PFC) added to University of Wisconsin (UW) and histidine-tryptofane-ketoglutarate (HTK) solutions on hepatocyte preservation. METHOD: Rat hepatocytes were isolated by colagenase digestion technique, with a viability greater than 85% measured by tripan blue exclusion. We suspended 1.2 x 10(8) hepatocytes in 20 mL of HTK solution, HTK with perfluor-n-octane (HTK + PFC), UW solution, and E. Williams medium as control group (CG). Five samples of each group were stored at 4 degrees C for 40 h. We measured Cell viability, large membrane bleb formation and extracellular LDH at 0, 12, 24 and 36 hours. RESULTS: Cell viability was lower in the UW at 12 h (p < 0.05) and 24 h (p < 0.05) compared to the rest of the study groups; at 36 h we found no differences in cell viability. There were fewer hepatocytes with large membrane blebs in UW + PFC; compared with the remainder of the solutions, but this difference was only reached statistical significance when compared to UW at 24 h (p < 0.05) and at 36 h (p < 0.05). When comparing groups with or without fluorocarbons LDH levels showed no difference at 0, 12, 24, and 36 h. CONCLUSIONS: The present study shows that the addition of fluorocarbons to UW solution diminishes large membrane formation (irreversible damage) and improves cell viability at 12 and 24 h. However, fluorocarbons added to both solutions failed to increase preservation time beyond 24 h.