Dutkowski P, Schönfeld S, Odermatt B, Heinrich T, Junginger T
Department of Surgery, University of Mainz, Germany.
Cryobiology. 1998 Feb;36(1):61-70. doi: 10.1006/cryo.1997.2066.
Rat livers were preserved hypothermically for 10 or 24 h in vitro as if for transplantation. Two methods of preservation were compared using physiological and biochemical parameters: simple storage and oscillating perfusion. By measuring the nucleotides after preservation the calculated energy charge was significantly higher after 10 and 24 h of oscillating perfusion compared to the simple storage group. In addition, a significant energy charge loading was demonstrated by 10 h oscillating perfusion compared to the initial value prior to perfusion. The oscillating, computer-controlled perfusion permits continuous monitoring of perfusate temperature, O2 consumption, pCO2, portal vein pressure, and pH and also automatic sample collection and pH compensation. In addition, the perfusate can be easily exchanged by using two different pumps or be rewarmed by a heat exchanger. For measuring of short-lived metabolites (interleukins, oxygen radicals, prostaglandins) sampling can be performed directly out of the vena cava outflow. pH and temperature stability was maintained by a data acquisition and controlling system. Because of a special designed liver chamber a combination of storage and perfusion with or without substrates was possible. The demonstrated standardized perfusion technique was achieved by a combination of special equipment and computer-aided monitoring and allows further experiments to improve understanding of ischemic and reperfusion injury.
大鼠肝脏在体外进行低温保存10或24小时，如同用于移植一样。使用生理和生化参数比较了两种保存方法：简单储存和振荡灌注。通过测量保存后的核苷酸，与简单储存组相比，振荡灌注10小时和24小时后计算出的能荷显著更高。此外，与灌注前的初始值相比，10小时振荡灌注显示出显著的能荷增加。振荡式计算机控制灌注允许连续监测灌注液温度、氧气消耗、pCO2、门静脉压力和pH值，还能自动进行样本采集和pH补偿。此外，通过使用两个不同的泵可以轻松更换灌注液，或者通过热交换器进行复温。为了测量短寿命代谢物（白细胞介素、氧自由基、前列腺素），可以直接从腔静脉流出物中进行采样。pH值和温度稳定性由数据采集和控制系统维持。由于特殊设计的肝脏腔室，有可能结合有或没有底物的储存和灌注。所展示的标准化灌注技术是通过特殊设备与计算机辅助监测相结合实现的，并且允许进行进一步的实验，以增进对缺血和再灌注损伤的理解。
