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pp 1-10 | Cite as

Isolation and Characterization of Extracellular Vesicles from Keratinocyte Cultures

  • Sebastian Sjöqvist
  • Aya Imafuku
  • Danu Ghupta
  • Samir E. L. Andaloussi
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Extracellular vesicles (EVs), including exosomes, are nano-sized membrane-bound particles which are released by cells. They have been found in all examined body fluids and can be isolated from conditioned cell culture media. These vesicles have gained increasing attention due to their importance in cellular cross talk, in both health and disease. For example, keratinocyte-derived EVs have been described to modulate melanin production in epidermis. Similar EVs were also shown to have an important role in skin immunology, by stimulating dendritic cells. In this chapter, we will describe how to isolate EVs from keratinocyte cultures and how to perform characterization by Western blot, nanoparticle tracking analysis, and transmission electron microscopy.

Keywords

Extracellular vesicles Exosomes Keratinocytes Epidermis EV isolation EV characterization 

Notes

Acknowledgments

This work was supported by the Swedish Society of Medicine. SELA is supported by the Swedish Research Council (VR-Med) and the Swedish foundation of Medical Research (SSF-IRC).

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Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Sebastian Sjöqvist
    • 1
  • Aya Imafuku
    • 2
  • Danu Ghupta
    • 3
  • Samir E. L. Andaloussi
    • 3
  1. 1.Department of Clinical SciencesIntervention and Technology (CLINTEC), Karolinska InstitutetStockholmSweden
  2. 2.Institute of Advanced Biomedical Engineering and ScienceTokyo Women’s Medical UniversityTokyoJapan
  3. 3.Department of Laboratory Medicine, Clinical Research CenterKarolinska InstitutetStockholmSweden

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