Differences in the lipid patterns during maturation of 3T3-L1 adipocytes investigated by thin-layer chromatography, gas chromatography, and mass spectrometric approaches

Anal Bioanal Chem. 2020 Apr;412(10):2237-2249. doi: 10.1007/s00216-019-02243-w. Epub 2019 Dec 3.

Abstract

Populations of industrialized countries have registered a dramatically increasing prevalence in obesity for many years. Despite continuous research, mechanisms involved in the storage and utilization of chemical energy in adipocytes are still under investigation. Adipocytes have the task to store excessive energy in the form of triacylglycerols (TG) and it is already well-known that the fatty acyl composition of TG is largely determined by the composition of the diet. In contrast to TG, the composition of adipocyte phospholipids was less comprehensively investigated. In this study, the compositions of the most abundant phospholipid classes of 3T3-L1 undifferentiated (preadipocytes) and differentiated cells (adipocytes) were determined. The lipid fractions were isolated by normal phase high-performance thin-layer chromatography and subsequently analyzed by electrospray ionization mass spectrometry. Additionally, the fatty acyl (FA) compositions were determined by gas chromatography. The positions of the FA residues were additionally confirmed by phospholipase A2 digestion. The advantages and disadvantages of the different analytical approaches will be discussed. It will be shown that undifferentiated 3T3-L1 and mature adipocytes differ extremely regarding their compositions. This goes along with an increase in odd-chain fatty acids. Graphical abstract.

Keywords: 3T3-L1 adipocytes; ESI mass spectrometry; Gas chromatography; Odd-chain fatty acids; Phospholipase A2 digestion; Thin-layer chromatography.

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / chemistry*
  • Adipocytes / cytology*
  • Adipocytes / metabolism
  • Animals
  • Cell Differentiation
  • Chromatography, Thin Layer / methods
  • Fatty Acids / chemistry
  • Fatty Acids / metabolism
  • Gas Chromatography-Mass Spectrometry / methods
  • Lipid Metabolism*
  • Lipids / chemistry*
  • Mice
  • Triglycerides / chemistry
  • Triglycerides / metabolism

Substances

  • Fatty Acids
  • Lipids
  • Triglycerides