Porretti L, Gatti S, Gramignoli R, Colombo F, Lopa R, Cattaneo A, Scalamogna M, Colombo G, Rossi G, Bonino F, Rebulla P, Prati D
Center of Transfusion Medicine, Cell Therapy and Cryobiology, IRCCS Fondazione Ospedale Policlinico, Mangiagalli e Regina Elena, Via Francesco Sforza 35, 20122 Milan, Italy.
Dig Liver Dis. 2006 Dec;38(12):905-11. doi: 10.1016/j.dld.2006.07.009. Epub 2006 Aug 21.
Although there is a growing interest on the use of non-heart beating donors to enlarge the liver donor pool, livers with prolonged warm ischaemia time are not currently considered for organ transplantation. We hypothesised that these organs may represent a source of hepatocytes for cell transplantation and/or use in bioartificial liver devices. Thus, we investigated if prolonged ischaemia could influence the recovery and viability of functional hepatocytes dissociated from rat livers.
Hepatocytes were isolated from the liver within 15 min after death (t=15 min) and after 4, 8 and 12h of ischaemia. Cells were either maintained in culture or cryopreserved. In all products, we evaluated cell recovery and viability, hepatocyte markers and cellular functions, including albumin and urea production.
The number of cells per gram of tissue was similar at 15 min, 4 and 8h, while it was significantly decreased at 12h. About 0.2 x 10(6) viable cells expressing hepatocyte markers and producing albumin and urea were isolated up to 8h of ischaemia per gram of tissue.
Recovery of viable and functional hepatocytes seems possible after prolonged ischaemia time. These data warrant the evaluation of hepatocyte isolation from human livers of non-heart beating donors.
尽管人们越来越关注利用非心脏跳动供体来扩大肝脏供体库，但目前器官移植不考虑热缺血时间延长的肝脏。我们推测这些器官可能是细胞移植和/或用于生物人工肝装置的肝细胞来源。因此，我们研究了长时间缺血是否会影响从大鼠肝脏分离的功能性肝细胞的恢复和活力。
在死亡后15分钟（t = 15分钟）以及缺血4、8和12小时后从肝脏中分离肝细胞。细胞要么进行培养，要么进行冷冻保存。在所有产物中，我们评估了细胞回收率和活力、肝细胞标志物以及细胞功能，包括白蛋白和尿素的产生。
每克组织的细胞数量在15分钟、4小时和8小时时相似，而在12小时时显著减少。每克组织在缺血长达8小时后可分离出约0.2×10⁶个表达肝细胞标志物并产生白蛋白和尿素的活细胞。
长时间缺血后似乎有可能恢复有活力和功能的肝细胞。这些数据值得对从非心脏跳动供体的人类肝脏中分离肝细胞进行评估。
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