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Complexity of fatty acid distribution inside human macrophages on single cell level using Raman micro-spectroscopy

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Abstract

Macrophages are phagocytic cells which are involved in the non-specific immune defense. Lipid uptake and storage behavior of macrophages also play a key role in the development of atherosclerotic lesions within walls of blood vessels. The allocation of exogenous lipids such as fatty acids in the blood stream dictates the accumulation and quantity of lipids within macrophages. In case of an overexposure, macrophages transform into foam cells because of the large amount of lipid droplets in the cytoplasm. Raman micro-spectroscopy is a powerful tool for studying single cells due to the combination of microscopic imaging with spectral information. With a spatial resolution restricted by the diffraction limit, it is possible to visualize lipid droplets within macrophages. With stable isotopic labeling of fatty acids with deuterium, the uptake and storage of exogenously provided fatty acids can be investigated. In this study, we present the results of time-dependent Raman spectroscopic imaging of single THP-1 macrophages incubated with deuterated arachidonic acid. The polyunsaturated fatty acid plays an important role in the cellular signaling pathway as being the precursor of icosanoids. We show that arachidonic acid is stored in lipid droplets but foam cell formation is less pronounced as with other fatty acids. The storage efficiency in lipid droplets is lower than in cells incubated with deuterated palmitic acid. We validate our results with gas chromatography and gain information on the relative content of arachidonic acid and its metabolites in treated macrophages. These analyses also provide evidence that significant amounts of the intracellular arachidonic acid is elongated to adrenic acid but is not metabolized any further. The co-supplementation of deuterated arachidonic acid and deuterated palmitic acid leads to a non-homogenous storage pattern in lipid droplets within single cells.

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Acknowledgments

We thank Carsten Rohrer for conducting the GC measurements, Maria Wallert and Lisa Schmölz for the introduction into the cell culture work and Dr. Iwan W. Schie for helpful discussions. We gratefully acknowledge the financial support by the Carl Zeiss Stiftung and the “Jenaer Biochip Initative 2.0” (JBCI 2.0). The project “JBCI 2.0” (03IPT513Y) within the framework “InnoProfile-Transfer–Unternehmen Region” is supported by the Federal Ministry of Education and Research (BMBF), Germany.

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Authors and Affiliations

  1. Leibniz Institute of Photonic Technology (IPHT), Albert-Einstein-Straße 9, 07745, Jena, Germany

    Clara Stiebing, Christian Matthäus, Christoph Krafft, Karina Weber & Jürgen Popp

  2. Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany

    Clara Stiebing, Christian Matthäus, Christoph Krafft, Karina Weber & Jürgen Popp

  3. Institute of Nutrition and Abbe Center of Photonics, Friedrich Schiller University Jena, Dornburger Straße 25, 07743, Jena, Germany

    Andrea-Anneliese Keller & Stefan Lorkowski

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  1. Clara Stiebing
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  2. Christian Matthäus
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  3. Christoph Krafft
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  4. Andrea-Anneliese Keller
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  6. Stefan Lorkowski
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  7. Jürgen Popp
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Correspondence to Jürgen Popp.

Additional information

Stefan Lorkowski and Jürgen Popp contributed equally to this work.

Published in the topical collection Single Cell Analysis with guest editors Petra Dittrich and Norbert Jakubowski.

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Stiebing, C., Matthäus, C., Krafft, C. et al. Complexity of fatty acid distribution inside human macrophages on single cell level using Raman micro-spectroscopy. Anal Bioanal Chem 406, 7037–7046 (2014). https://doi.org/10.1007/s00216-014-7927-0

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  • DOI: https://doi.org/10.1007/s00216-014-7927-0

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