Laboratory of Liver Research, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea.
Department of Internal Medicine, Chungnam National University School of Medicine, Daejeon, Korea.
Mol Cells. 2019 Jan 31;42(1):45-55. doi: 10.14348/molcells.2018.0330. Epub 2018 Dec 12.
The liver is involved in a wide range of activities in vertebrates and some other animals, including metabolism, protein synthesis, detoxification, and the immune system. Until now, various methods have been devised to study liver diseases; however, each method has its own limitations. liver perfusion machinery, originally developed in rats, has been successfully adapted to mice, enabling the study of liver diseases. Here we describe the protocol, which is a simple but widely applicable method for investigating the liver diseases. The liver is perfused by cannulation of the portal vein and suprahepatic inferior vena cava (IVC), with antegrade closed circuit circulation completed by clamping the infrahepatic IVC. liver perfusion can be utilized to evaluate immune cell migration and function, hemodynamics and related cellular reactions in each type of hepatic cells, and the metabolism of toxic or other compounds by changing the composition of the circulating media. liver perfusion method maintains liver function and cell viability for up to 2 h. This study also describes an optional protocol using density-gradient centrifugation for the separation of different types of hepatic cells, allowing the determination of changes in each cell type. In summary, this method of liver perfusion will be useful for studying liver diseases as a complement to other established methods.
肝脏在脊椎动物和其他一些动物中参与广泛的活动,包括新陈代谢、蛋白质合成、解毒和免疫系统。到目前为止,已经设计了各种方法来研究肝脏疾病;然而,每种方法都有其自身的局限性。肝脏灌流设备最初是在大鼠中开发的,现已成功适应于小鼠,使肝脏疾病的研究成为可能。在这里,我们描述了一种方案,这是一种简单但广泛适用的研究肝脏疾病的方法。通过门静脉和肝上腔下腔静脉(IVC)的插管进行肝脏灌流,通过夹闭肝下 IVC 完成顺行闭路循环。通过改变循环介质的组成,可以利用肝脏灌流来评估免疫细胞的迁移和功能、肝内各类型细胞的血液动力学和相关细胞反应以及有毒或其他化合物的代谢。肝脏灌流方法可维持肝脏功能和细胞活力长达 2 小时。本研究还描述了一种可选方案,使用密度梯度离心分离不同类型的肝细胞,从而可以确定每种细胞类型的变化。总之,这种肝脏灌流方法将有助于作为其他已建立方法的补充来研究肝脏疾病。