Direct visualization of extracellular vesicles on the membrane of human mesenchymal stem/stromal cells by cryo-electron microscopy

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Abstract

Extracellular vesicles (EVs) play an important role in intercellular communication and influence a wide range of physiological and pathological processes. Membrane-associated extracellular vesicles (MAVs) represent a distinct and poorly understood class of EVs. This study demonstrates the application of cryo-electron microscopy (cryo-EM) to investigate MAVs secreted by human mesenchymal stem/stromal cells (MSCs). Cryo-EM revealed vesicles ranging in diameter from 50 to 750 nm located near the cell surface. The results obtained will facilitate further studies on the physiological role of MAVs and their association with cell membranes.

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About the authors

A. V. Moiseenko

Biological Department of Lomonosov

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0003-1112-2356

Researcher

Russian Federation, Moscow

N. A. Basalova

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0002-2597-8879

Cand. of Sci. (Biology), Junior Research Assistant

Russian Federation, Moscow

D. V. Bagrov

Biological Department of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0002-6355-7282

Cand. of Sci. (Physics and Mathematics), Leading Researcher

Russian Federation, Moscow

T. S. Trifonova

Biological Department of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0003-2042-5244

Laboratory assistant

Russian Federation, Moscow

M. A. Vigovsky

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0003-2103-8158

Laboratory assistant

Russian Federation, Moscow

U. D. Dyachkova

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0002-6119-8976

Laboratory assistant

Russian Federation, Moscow

O. A. Grigorieva

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0003-2954-2420

Cand. of Sci. (Biology)

Russian Federation, Moscow

E. S. Novoseletskaya

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0002-0922-9157

Cand. of Sci. (Biology)

Russian Federation, Moscow

A. Y. Efimenko

Medical Research and Education Institute of Lomonosov MSU

Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0002-0696-1369

Doct. of Sci. (Physiology), Head of Laboratory

Russian Federation, Moscow

O. S. Sokolova

Biological Department of Lomonosov MSU

Author for correspondence.
Email: sokolova@mail.bio.msu.ru
ORCID iD: 0000-0003-4678-232X

Doct. of Sci. (Biology), Prof.

Russian Federation, Moscow

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2. Fig.1. Photographs of MSCs on a gold grid: а – photograph obtained during cell culture using light microscopy (scale bar is 50 μm), the purple arrows indicate the cell edges, b – cryo EM image of a cell frozen on a perforated carbon substrate (scale bar is 5 μm)

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3. Fig.2. Micrograph of the MSC edge with vesicles located near it, obtained by cryo EM method (scale segment is 200 nm). In image a the contrast is increased for clarity. In image b, individual vesicles and cytoskeleton elements are highlighted in colour: white – ice contamination, blue – lipid bilayers of EVs and cell edge, orange – hole in carbon substrate (diameter is 1.2 μm), pink – cell cytoskeleton

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4. Fig.3. Micrograph of multilayer EVs near the MSC cell edge obtained by cryo EM (scale segment is 200 nm). In image a the contrast is increased for clarity. In image b individual vesicles, cell edge and cytoskeleton elements are highlighted in colour: blue – lipid bilayers of MSC cell and EVs, pink – cytoskeleton

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Copyright (c) 2024 Moiseenko A.V., Basalova N.A., Bagrov D.V., Trifonova T.S., Vigovsky M.A., Dyachkova U.D., Grigorieva O.A., Novoseletskaya E.S., Efimenko A.Y., Sokolova O.S.