Terry Claire, Hughes Robin D
Institute of Liver Studies, King's College London School of Medicine London, UK.
Methods Mol Biol. 2009;481:25-34. doi: 10.1007/978-1-59745-201-4_3.
Successful cryopreservation of hepatocytes is essential for their use in hepatocyte transplantation. Cryopreservation allows hepatocytes to be available for emergency treatment of acute liver failure and also for planned treatment of liver-based metabolic disorders. In addition, cryopreservation of human hepatocytes can facilitate their use in metabolism and toxicity studies. Cryopreservation can adversely affect the viability and function, especially reduce the attachment efficiency, of hepatocytes on thawing.The cryopreservation process can be divided into steps so that improvements can be made on the 'standard' protocols that are followed in some laboratories. These steps are as follows: pre-incubation of cells; freezing solution, cryoprotectants and cytoprotectants; freezing process; storage; thawing; post-thawing culture. This chapter presents an optimised protocol for cryopreservation of human hepatocytes as developed at King's College Hospital.
成功冻存肝细胞对于其在肝细胞移植中的应用至关重要。冻存使得肝细胞可用于急性肝衰竭的紧急治疗以及基于肝脏的代谢紊乱的计划性治疗。此外，人肝细胞的冻存有助于其在代谢和毒性研究中的应用。冻存会对肝细胞解冻后的活力和功能产生不利影响，尤其是降低其贴壁效率。冻存过程可分为多个步骤，以便对一些实验室所遵循的“标准”方案进行改进。这些步骤如下：细胞预孵育；冷冻溶液、冷冻保护剂和细胞保护剂；冷冻过程；储存；解冻；解冻后培养。本章介绍了国王学院医院制定的人肝细胞冻存优化方案。
