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 to investigate whether the addition of SAMe to EuroCollins solution (EC) could provide a similar degree of protection as the more complex University of Wisconsin (UW) solution during cold preservation. In addition, we have investigated a possible protective action of SAMe during hepatocyte cryopreservation. Wistar rat hepatocytes (10(6) cells/ml) were stored in either EC (+/- 12 mumol/l SAMe) or UW. In parallel, hepatocytes (10(6) cells/ml) were cryopreserved in M199 culture medium (+/- SAMe) using dimethyl sulfoxide as cryoprotectant. LDH release, viability, and hepatocyte GSH and malondialdehyde (MDA) content were sequentially determined during cold preservation. There were no differences on viability or GSH and MDA content between EC+SAMe and UW stored cells, although LDH release was slightly higher in the first group. The addition 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), observed after thawing. Our results suggest that SAMe could be a useful additive for both cold storage and cryopreservation solutions of hepatocytes.
Vara
E
E
Department of Biochemistry (Facultad de Medicina), Universidad Complutense, Madrid, Spain.
Arias-Díaz
J
J
Villa
N
N
Hernández
J
J
García
C
C
Ortiz
P
P
Balibrea
J L
JL
D000818:Animals; D002452:Cell Count; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D005978:Glutathione / Q000378:metabolism; D006982:Hypertonic Solutions; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000494:pharmacology*; D013997:Time Factors
C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D017382:Reactive Oxygen Species; D008315:Malondialdehyde; D012436:S-Adenosylmethionine; D007770:L-Lactate Dehydrogenase; D005978:Glutathione