The effect of antioxidants and a caspase inhibitor on cryopreserved rat hepatocytes.
Hepatocyte transplantation and use of bioartificial liver support systems have been suggested as potential therapies for fulminant hepatic failure. Cryopreservation in liquid nitrogen is presently the major method of long-term storage of isolated hepatocytes. However, cryopreservation can result in low cell recovery and reduction in differentiated function. Several possible mechanisms of cell death during cryopreservation have been proposed. The most important mechanisms appear to be oxidative stress and apoptosis. In this study, we isolated fresh rat hepatocytes and cryopreserved them in three media: University of Wisconsin (UW) solution, an antioxidant-containing medium, and medium containing a caspase inhibitor. Viability and function of hepatocytes cryopreserved in these media were examined. Cryopreservation conditions had no effect on hepatocyte viability after thawing. However, after culture we found significant improvements in viability and function in both antioxidant- and caspase inhibitor-treated hepatocytes at 6 and 24 h.
Fujita
Rie
R
Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Hui
Thomas
T
Chelly
Marjorie
M
Demetriou
Achilles A
AA
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D003470:Culture Media; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D019164:Liver, Artificial; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats
D000975:Antioxidants; D061945:Caspase Inhibitors; D003470:Culture Media; D011480:Protease Inhibitors; D020169:Caspases