An in vitro model for essential fatty acid deficiency: HepG2 cells permanently maintained in lipid-free medium

E. E. Furth, H. Sprecher, E. A. Fisher, H. D. Fleishman, M. Laposata

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Abstract

A stable essential fatty acid-deficient cell type, known as HepG2-EFD, was derived from the lipoprotein-producing human hepatoma cell line HepG2. These cells are particularly useful for quantitative studies involving essential fatty acids (n-6 and n-3 fatty acids) in secreted lipoproteins. Radiolabeled essential fatty acids can be delivered to these cells without altering the specific activity of the fatty acids, since the deficient cells contain no endogenous essential fatty acids. Using these cells, radioactivity data (dpm) from metabolic studies can be converted directly to mass, and masses as low as a few pmoles can be accurately measured. HepG2-EFD cell cultures were established by growing HepG2 cells in medium containing delipidated serum. After 10 days of growth in delipidated medium, HepG2 cells were completely depleted of all essential fatty acids. Compensatory increases in nonessential fatty acids (n-9 and n-7 fatty acids) including 20:3n-9 (the Mead acid), which is the hallmark fatty acid of essential fatty acid deficiency, were also observed in HepG2-EFD cells. Despite the lack of exogenous fatty acids in the medium and the lack of essential fatty acids in the cells, export of very low density lipoprotein (VLDL)-associated apolipoprotein B by HepG2-EFD was the same as observed for parent HepG2 cells. However, the activity of beta-oxidation of fatty acids in HepG2-EFD cells was much lower than in the parent cell line. The doubling time was the same for both cell types, but depletion of essential fatty acids resulted in a phenotypic change, whereby HepG2 cells proliferated in large clusters of cells, but HepG2-EFD cells proliferated as homogeneously dispersed cell monolayers. Thus, cultures of HepG2-EFD cells provide an in vitro model for essential fatty acid deficiency, and the model has fatty acid changes consistent with those found in vivo in essential fatty acid deficiency.

Original languageEnglish (US)
Pages (from-to)1719-1726
Number of pages8
JournalJournal of Lipid Research
Volume33
Issue number11
StatePublished - Jan 1 1992
Externally publishedYes

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Keywords

  • Mead acid
  • arachidonate
  • essential fatty acids

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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