Guillouzo A, Rialland L, Fautrel A, Guyomard C
INSERM U 456, Faculté de Pharmacie, Université de Rennes, France.
Chem Biol Interact. 1999 Jun 1;121(1):7-16. doi: 10.1016/s0009-2797(99)00087-3.
Cryopreservation in liquid nitrogen is presently the only way for long-term storage of isolated hepatocytes. Freeze-thaw conditions are not well defined yet; the most critical parameters appear to be the choice of the cryoprotectant, composition of the freezing medium, and cooling and thawing rates. Comparable results have been obtained with hepatocytes from various species, including man. Cryopreservation usually results in low cell recovery and early alterations of functional activities. However, both phase I and phase II xenobiotic metabolism is still active after thawing, at least during a short period. Moreover, survival and function of cryopreserved hepatocytes can be improved when these cells have a high energy status, are cryopreserved after immobilization in a gel, separated from dead cells on a Percoll gradient or placed in more favorable culture conditions (e.g. in coculture with liver non parenchymal cells). Additional studies are needed to improve freeze-thaw protocols and to better characterize liver parenchymal cells after storage, including evaluation of their responsiveness to specific inducers.
液氮冷冻保存是目前长期储存分离的肝细胞的唯一方法。冻融条件尚未明确界定；最关键的参数似乎是冷冻保护剂的选择、冷冻培养基的组成以及冷却和解冻速率。来自包括人类在内的各种物种的肝细胞都获得了类似的结果。冷冻保存通常会导致细胞回收率低和功能活性的早期改变。然而，解冻后，至少在短时间内，I相和II相外源性物质代谢仍然活跃。此外，当这些细胞具有高能量状态、在固定于凝胶中后进行冷冻保存、通过Percoll梯度与死细胞分离或置于更有利的培养条件下（例如与肝非实质细胞共培养）时，冷冻保存的肝细胞的存活率和功能可以得到改善。需要进行更多的研究来改进冻融方案，并更好地表征储存后的肝实质细胞，包括评估它们对特定诱导剂的反应性。
