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

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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.

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Acknowledgments

We thank Prof. Dr. Ines Liebscher, Dr. Doreen Thor, and Tomáš Suchý for providing 3T3-L1 cells and the relevant microscopic images.

Funding

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.

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Authors and Affiliations

  1. Institute for Medical Physics and Biophysics, Medical Faculty, Leipzig University, Härtelstr. 16-18, 04107, Leipzig, Germany

    Yulia Popkova, Jürgen Schiller & Kathrin M. Engel

  2. Leibniz Institute for Farm Animal Biology, Institute of Muscle Biology and Growth, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Dirk Dannenberger

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  1. Yulia Popkova
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Correspondence to Yulia Popkova.

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Published in the topical collection Current Progress in Lipidomics with guest editors Michal Holčapek, Gerhard Liebisch, and Kim Ekroos.

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Popkova, Y., Dannenberger, D., Schiller, J. et al. 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 412, 2237–2249 (2020). https://doi.org/10.1007/s00216-019-02243-w

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