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Updated: Jul 19, 2025
Cholesterol Efflux Assay
Published on: March 6, 2012
Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic
Jonathan D Cybulski1, Kit Sum Leung2, Carmen Oi Ning Leung3
1School of Biological Sciences, Faculty of Science, The University of Hong Kong, Hong Kong, Hong Kong SAR; Smithsonian Tropical Research Institute, Balboa, Republic of Panama.
Abstract:
Cholesterol biosynthesis supports proliferation and drives resistance to tyrosine kinase inhibitor (TKI) therapy in hepatocellular carcinoma (HCC). Here, we present a protocol for using stable isotopic tracers to track the biosynthesis of cholesterol in cultured HCC cells. We describe steps for cell preparation, incubation, separation, and homogenization. We then detail lipid extraction and compound-specific isotope analysis for comparing and quantifying cholesterol synthesis between TKI-resistant HCC cells and their mock counterparts. This protocol can be expanded for use with other shorter-chained lipids.
