4.5 Article

Microscopic characterization reveals the diversity of EVs secreted by GFP-HAS3 expressing MCF7 cells

Journal

EUROPEAN JOURNAL OF CELL BIOLOGY
Volume 101, Issue 3, Pages -

Publisher

ELSEVIER GMBH
DOI: 10.1016/j.ejcb.2022.151235

Keywords

Extracellular vesicle; Morphology; Hyaluronan synthase; Confocal microscopy; 3D culture; Electron tomography

Categories

Funding

  1. Academy of Finland GeneCellNano Flagship [337120]
  2. Jane and Aatos Erkko Foundation
  3. Mizutani Foundation
  4. Japan Paavo Koistinen Foundation
  5. Northern Savo Cancer Foundation
  6. EATRIS
  7. European Infrastructure for Translational Medicine

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We used high-resolution imaging techniques to characterize the size and morphology of extracellular vesicles produced by breast cancer cells. The results showed diverse morphology of these vesicles and emphasized the importance of comparing different methods.
We have shown the connection of hyaluronan synthesis activity with the enhanced shedding of extracellular vesicles, but detailed morphological analysis of those hyaluronan-induced EVs is still missing. In this study we utilized a comprehensive set of high-resolution imaging techniques to characterize in high detail the size and morphology of EVs originating from stable MCF7 breast cancer cell line and transiently transfected cells expressing GFP-HAS3. To avoid possible artefacts or loss of EVs resulting from the isolation process, special attention was paid to analysis of EVs in situ in monolayer and in 3D cultures. The results of this study show that GFP-HAS3 expressing MCF7 cells produce morphologically diverse EVs but also demonstrates the variation in results obtained with different experimental setup, which emphasizes the importance of comparison between different methods when interpreting the observations.

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