Department of Medicine, Division of Cardiovascular Medicine
Cellular and Molecular Physiology | Molecular Biology
Lipid metabolism in liver is complex. In addition to importing and exporting lipid via lipoproteins, hepatocytes can oxidize lipid via fatty acid oxidation, or alternatively, synthesize new lipid via de novo lipogenesis. The net sum of these pathways is dictated by a number of factors, which in certain disease states leads to fatty liver disease. Excess hepatic lipid accumulation is associated with whole body insulin resistance and coronary heart disease. Tools to study lipid metabolism in hepatocytes are useful to understand the role of hepatic lipid metabolism in certain metabolic disorders. In the liver, hepatocytes regulate the breakdown and synthesis of fatty acids via beta-fatty oxidation and de novo lipogenesis, respectively. Quantifying metabolism in these pathways provides insight into hepatic lipid handling. Unlike in vitro quantification, using primary hepatocytes, making measurements in vivo is technically challenging and resource intensive. Hence, quantifying beta-fatty acid oxidation and de novo lipogenesis in cultured mouse hepatocytes provides a straight forward method to assess hepatocyte lipid handling. Here we describe a method for the isolation of primary mouse hepatocytes, and we demonstrate quantification of beta-fatty acid oxidation and de novo lipogenesis, using radiolabeled substrates.
Molecular Biology, Issue 102, Liver, Hepatocyte, Mouse, Fatty Acid, Oxidation, Lipogenesis, Metabolism, Palmitate
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DOI of Published Version
J Vis Exp. 2015 Aug 27;(102):e52982. doi: 10.3791/52982. Link to article on publisher's site
Journal of visualized experiments : JoVE
Akie TE, Cooper MP. (2015). Determination of Fatty Acid Oxidation and Lipogenesis in Mouse Primary Hepatocytes. GSBS Student Publications. https://doi.org/10.3791/52982. Retrieved from https://escholarship.umassmed.edu/gsbs_sp/2004