Differences in the lipid patterns during maturation of 3T3-L1 adipocytes investigated by thin-layer chromatography, gas chromatography, and mass spectrometric approaches
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.
Keywords3T3-L1 adipocytes Odd-chain fatty acids ESI mass spectrometry Thin-layer chromatography Gas chromatography Phospholipase A2 digestion
We thank Prof. Dr. Ines Liebscher, Dr. Doreen Thor, and Tomáš Suchý for providing 3T3-L1 cells and the relevant microscopic images.
This study was supported by the German Research Council (SFB 1052/Z3 (Project Number 209933838)). We are particularly indebted to the MERCK Company for the continuous support.
Compliance with ethical standards
Conflict of Interest
The authors declare that they have no conflict of interest.
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