Mol. Cells 2018; 41(): 953  https://doi.org/10.14348/molcells.2018.0330
Experimental Applications of in situ Liver Perfusion Machinery for the Study of Liver Disease
Won-Mook Choi1, Hyuk Soo Eun2, Young-Sun Lee3, Sun Jun Kim2, Myung-Ho Kim1, Jun-Hee Lee1, Young-Ri Shim1, Hee-Hoon Kim1, Ye Eun Kim1, Hyon-Seung Yi1,2,*, and Won-Il Jeong1,*
1Laboratory of Liver Research, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea, 2Department of Internal Medicine, Chungnam National University School of Medicine, Daejeon, Korea, 3Department of Internal Medicine, Korea University College of Medicine, Korea
*Correspondence: jmpbooks@cnuh.co.kr (HSY); wijeong@kaist.ac.kr (WIJ)
Received August 2, 2018; Revised September 29, 2018; Accepted October 18, 2018.; Published online December 12, 2018.
© Korean Society for Molecular and Cellular Biology. All rights reserved.

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ABSTRACT
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. In situ 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 in situ by cannulation of the portal vein and suprahepatic inferior vena cava (IVC), with antegrade closed circuit circulation completed by clamping the infrahepatic IVC. In situ 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. In situ liver perfusion method maintains liver function and cell viability for up to 2 h.
This study also describes an optional protocol using densitygradient centrifugation for the separation of different types of hepatic cells, allowing the determination of changes in each cell type. In summary, this method of in situ liver perfusion will be useful for studying liver diseases as a complement to other established methods.
Keywords: hemodynamics, immune cell, in situ perfusion, liver disease, metabolism


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