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Cell density-induced changes in lipid composition and intracellular trafficking

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Abstract

Cell density is one of the extrinsic factors to which cells adapt their physiology when grown in culture. However, little is known about the molecular changes which occur during cell growth and how cellular responses are then modulated. In many cases, inhibitors, drugs or growth factors used for in vitro studies change the rate of cell proliferation, resulting in different cell densities in control and treated samples. Therefore, for a comprehensive data analysis, it is essential to understand the implications of cell density on the molecular level. In this study, we have investigated how lipid composition changes during cell growth, and the consequences it has for transport of Shiga toxin. By quantifying 308 individual lipid species from 17 different lipid classes, we have found that the levels and species distribution of several lipids change during cell growth, with the major changes observed for diacylglycerols, phosphatidic acids, cholesterol esters, and lysophosphatidylethanolamines. In addition, there is a reduced binding and retrograde transport of Shiga toxin in high density cells which lead to reduced intoxication by the toxin. In conclusion, our data provide novel information on how lipid composition changes during cell growth in culture, and how these changes can modulate intracellular trafficking.

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Acknowledgments

The work performed by the Oslo group has been supported by The Norwegian Council for Science and Humanities, The Norwegian Cancer Society, and Southern and Eastern Norway Regional Authority. The authors thank Anne Grethe Myrann for technical assistance with cell studies, Ellen Skarpen for assistance with microcopy studies, and Sirpa Sutela-Tuominen for assistance with the lipidomic analyzes.

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Authors and Affiliations

  1. Department of Biochemistry, Institute for Cancer Research, Oslo University Hospital, The Norwegian Radium Hospital, Montebello, 0379, Oslo, Norway

    Simona Kavaliauskiene, Carl-Martin Nymark, Jonas Bergan, Roger Simm, Tore Skotland & Kirsten Sandvig

  2. Center for Cancer Biomedicine, Oslo University Hospital, Oslo, Norway

    Simona Kavaliauskiene, Carl-Martin Nymark, Jonas Bergan, Roger Simm, Tore Skotland & Kirsten Sandvig

  3. Department of Biosciences, University of Oslo, Oslo, Norway

    Simona Kavaliauskiene, Jonas Bergan & Kirsten Sandvig

  4. Zora Biosciences, Espoo, Finland

    Tuulia Sylvänne, Helena Simolin & Kim Ekroos

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  1. Simona Kavaliauskiene
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  2. Carl-Martin Nymark
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Correspondence to Kirsten Sandvig.

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Kavaliauskiene, S., Nymark, CM., Bergan, J. et al. Cell density-induced changes in lipid composition and intracellular trafficking. Cell. Mol. Life Sci. 71, 1097–1116 (2014). https://doi.org/10.1007/s00018-013-1441-y

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