BENEFICIAL EFFECT OF S-ADENOSYLMETHIONINE DURING BOTH COLD-STORAGE AND CRYOPRESERVATION OF ISOLATED HEPATOCYTES

Oxygen free radicals appear to be the prime cell-toxic products during cold preservation. Glutathione (GSH) seems to play a critical role in cell protection against oxidant stress. The experimental decrease of intracellular GSH in vivo may be prevented by the administration of S- adenosylmethionine (SAMe), which seems also to play an important role in preserving the structure of cell membranes. We designed our study t o investigate whether the addition of SAMe to EuroCollins solution (EC ) could provide a similar degree of protection as the more complex Uni versity of Wisconsin (UW) solution during cold preservation. In additi on, we have investigated a possible protective action of SAMe during h epatocyte cryopreservation. Wistar rat hepatocytes (10(6) cells/ml) we re stored in either EC (+/-12 mu mol/l SAMe) or UW. In parallel, hepat ocytes (10(6) cells/ml) were cryopreserved in M199 culture medium (+/- SAMe) using dimethyl sulfoxide as cryoprotectant. LDH release, viabili ty, and hepatocyte GSH and malondialdehyde (MDA) content were sequenti ally determined during cold preservation. There were no differences on viability or GSH and MDA content between EC + SAMe and UW stored cell s, although LDH release was slightly higher in the first group. The ad dition of SAMe also attenuated the decrease in both viability (37 +/- 0.8 vs 53.0 +/- 7.4%, mean +/- SEM, N = 5, P < 0.05) and GSH content ( 13.4 +/- 15.1 vs 45.1 +/- 16.8%, mean +/- SEM, N = 5, P < 0.01), obser ved after thawing. Our results suggest that SAMe could be a useful add itive for both cold storage and cryopreservation solutions of hepatocy tes. (C) 1995 Academic Press, Inc.