Ferrigno Andrea, Richelmi Plinio, Vairetti Mariapia
Department of Internal Medicine and Therapeutics, University of Pavia, Italy.
J Pharmacol Toxicol Methods. 2013 Mar-Apr;67(2):107-14. doi: 10.1016/j.vascn.2012.10.001. Epub 2012 Oct 16.
Isolated perfused liver (IPL) model is not only widely performed in rats but is also used in mouse liver, although a detailed description of this procedure is absent. A comparison of the different techniques used on rats and mice will be discussed in this article association with a detailed description of the surgical and technical aspects needed to obtain and maintain the integrity of the livers during the organ isolation and perfusion.
The surgical procedures, the IPL set-up, and the evaluation of hepatic function and damage will be described in relation to both rats and mice. In particular, the heparin dosage and administration, the portal vein cannulation avoiding portal leakage, the use of suprahepatic caval vein output, and the insertion of a cannula for bile collection will be reported. For the settings, the perfusion circuit, the perfusion solution, the temperature and the flow rate will be described, with particular regard to the balance between perfusion pressure and oxygen delivery. The monitoring of liver integrity by measuring oxygen concentration and calculating oxygen delivery rate and oxygen uptake rate, and recommendations for the collection of perfusate and bile samples will be considered. Accurate pH measurement with normalization, and the perfusion portal pressure assay by a calibrated water manometer will be also reported.
This work analyzes the parameters crucial to performing a correct IPL both in rat and mouse, comparing our experience with the equivalent practice from other laboratories. An updated example of IPL applications in liver toxicology and pharmacology, physiology and pathophysiology, and liver graft preservation will be briefly presented, underlining how this technique provides essential information allowing a more accurate planning of the in vivo studies.
孤立灌注肝（IPL）模型不仅在大鼠中广泛应用，也用于小鼠肝脏，尽管缺乏对此操作的详细描述。本文将讨论在大鼠和小鼠上使用的不同技术的比较，并详细描述在器官分离和灌注过程中获取和维持肝脏完整性所需的手术和技术方面。
将描述与大鼠和小鼠相关的手术程序、IPL设置以及肝功能和损伤的评估。特别将报告肝素剂量和给药方式、避免门静脉漏血的门静脉插管、肝上腔静脉输出的使用以及胆汁收集插管的插入。对于设置，将描述灌注回路、灌注液、温度和流速，尤其关注灌注压力和氧气输送之间的平衡。将考虑通过测量氧气浓度和计算氧气输送率及氧气摄取率来监测肝脏完整性，以及收集灌注液和胆汁样本的建议。还将报告进行精确pH测量并进行标准化，以及使用校准的水柱压力计测量灌注门静脉压力。
这项工作分析了在大鼠和小鼠中进行正确IPL的关键参数，将我们的经验与其他实验室的等效实践进行了比较。将简要介绍IPL在肝脏毒理学、药理学、生理学和病理生理学以及肝移植保存中的最新应用实例，强调该技术如何提供重要信息，以便更准确地规划体内研究。
