Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease

Summary: We present a protocol for isolating hepatocytes from mice with established non-alcoholic fatty liver disease. This protocol consists of liver perfusion using a peristaltic pump, followed by a modified 25% and 90% Percoll gradient centrifugation protocol to capture lipid-laden hepatocytes th...

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Main Authors:	Yunshin Jung, Meng Zhao, Katrin J. Svensson
Format:	Article
Language:	English
Published:	Elsevier 2020-12-01
Series:	STAR Protocols
Subjects:	Cell Biology Cell culture Cell isolation Single Cell Metabolism Model Organisms
Online Access:	http://www.sciencedirect.com/science/article/pii/S2666166720302094

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spelling	doaj-c0375322196548eb89572dd5f9f646f12020-12-21T04:49:14ZengElsevierSTAR Protocols2666-16672020-12-0113100222Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver diseaseYunshin Jung0Meng Zhao1Katrin J. Svensson2Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USADepartment of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USADepartment of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USA; Corresponding authorSummary: We present a protocol for isolating hepatocytes from mice with established non-alcoholic fatty liver disease. This protocol consists of liver perfusion using a peristaltic pump, followed by a modified 25% and 90% Percoll gradient centrifugation protocol to capture lipid-laden hepatocytes that are usually lost using traditional isolation protocols. This protocol enables simultaneous isolation of normal and lipid-filled hepatocytes. Lipid-filled hepatocytes can be used in cell culture systems to study drug metabolism, hepatotoxicity, or glucose and lipid metabolism.For complete details on the use and execution of this protocol, please refer to Sharabi et al. (2017) and Kegel et al. (2016).http://www.sciencedirect.com/science/article/pii/S2666166720302094Cell BiologyCell cultureCell isolationSingle CellMetabolismModel Organisms
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Yunshin Jung Meng Zhao Katrin J. Svensson
spellingShingle	Yunshin Jung Meng Zhao Katrin J. Svensson Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease STAR Protocols Cell Biology Cell culture Cell isolation Single Cell Metabolism Model Organisms
author_facet	Yunshin Jung Meng Zhao Katrin J. Svensson
author_sort	Yunshin Jung
title	Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
title_short	Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
title_full	Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
title_fullStr	Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
title_full_unstemmed	Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
title_sort	isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
publisher	Elsevier
series	STAR Protocols
issn	2666-1667
publishDate	2020-12-01
description	Summary: We present a protocol for isolating hepatocytes from mice with established non-alcoholic fatty liver disease. This protocol consists of liver perfusion using a peristaltic pump, followed by a modified 25% and 90% Percoll gradient centrifugation protocol to capture lipid-laden hepatocytes that are usually lost using traditional isolation protocols. This protocol enables simultaneous isolation of normal and lipid-filled hepatocytes. Lipid-filled hepatocytes can be used in cell culture systems to study drug metabolism, hepatotoxicity, or glucose and lipid metabolism.For complete details on the use and execution of this protocol, please refer to Sharabi et al. (2017) and Kegel et al. (2016).
topic	Cell Biology Cell culture Cell isolation Single Cell Metabolism Model Organisms
url	http://www.sciencedirect.com/science/article/pii/S2666166720302094
work_keys_str_mv	AT yunshinjung isolationcultureandfunctionalanalysisofhepatocytesfrommicewithfattyliverdisease AT mengzhao isolationcultureandfunctionalanalysisofhepatocytesfrommicewithfattyliverdisease AT katrinjsvensson isolationcultureandfunctionalanalysisofhepatocytesfrommicewithfattyliverdisease
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Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease

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