Cytochrome P-450 content and activity after cold storage of rat hepatocytes in University of Wisconsin and sodium-lactobionate-sucrose solutions

We compared the capacity of University of Wisconsin (UW) and of sodium-lact obionate-sucrose (SLS) hypothermic preservation solutions to maintain the i ntegrity of the hepatic cytochrome P-450-dependent mono-oxygenase system. I solated rat hepatocytes were stored for 0, 10, 24, and 48 hours in UW or SL S solution and were subsequently cultured shortly at 37 degrees C, Cell via bility declined slightly but significantly in a time-dependent manner durin g cold preservation in either UW or SLS solution, and warm culture exacerba ted this effect, Total cytochrome P-450 declined gradually after cold prese rvation and warm culture to reach values of 70% and 52% of unstored control s in cells preserved for 24 and 48 hours in cold UW solution, respectively, Storage in cold SLS solution yielded a similar decrease to 79% and 59% of unstored controls for the equivalent preservation times, Cytochrome P-450 a ctivity was assessed by the metabolism of theophylline after Various cold p reservation times in UW or SLS solutions, Production of the major metabolit e 1,3-dimethyluric acid was not significantly affected by extended cold pre servation periods in either UW or SLS solutions. Similarly, the amount of r esidual theophylline remained stable in all groups, suggesting that alterna tive metabolic routes were not modified, These studies show that cold prese rvation in SLS solution is as effective as that in UW solution in terms of cell viability, cytochrome P-450 content, and activity toward theophylline. In addition, the significant reduction in cytochrome P-450 in conjunction with unaffected theophylline disposition suggests that certain cytochrome P -450 isoforms are specifically damaged by cold preservation and rewarming, Copyright (C) 1999 by the American Association for the Study of Liver Disea ses.