Fujita Rie, Hui Thomas, Chelly Marjorie, Demetriou Achilles A
Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Cell Transplant. 2005;14(6):391-6. doi: 10.3727/000000005783982981.
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.
肝细胞移植和生物人工肝支持系统的应用已被提议作为暴发性肝衰竭的潜在治疗方法。液氮冷冻保存目前是分离肝细胞长期储存的主要方法。然而，冷冻保存可能导致细胞回收率低和分化功能降低。已经提出了冷冻保存期间细胞死亡的几种可能机制。最重要的机制似乎是氧化应激和细胞凋亡。在本研究中，我们分离了新鲜大鼠肝细胞，并将它们保存在三种培养基中：威斯康星大学（UW）溶液、含抗氧化剂的培养基和含半胱天冬酶抑制剂的培养基。检测了在这些培养基中冷冻保存的肝细胞的活力和功能。冷冻保存条件对解冻后肝细胞活力没有影响。然而，培养后我们发现，在6小时和24小时时，抗氧化剂处理组和半胱天冬酶抑制剂处理组的肝细胞在活力和功能方面均有显著改善。
