Methodological Approaches to Assessing the Size and Morphology of Microvesicles of Cell Lines
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Cell Technologies in Biology and Medicine, No. 2, August, 2020
Methodological Approaches to Assessing the Size and Morphology of Microvesicles of Cell Lines
K. L. Markova1, A. R. Kozyreva1, A. A. Gorshkova1, E. P. Aleksandrova1, M. E. Berezkina1, V. A. Mikhailova1, A. N. Ivanova2, S. Yu. Kaputkina3, K. V. Onokhin1, K. A. Benken4, S. A. Sel’kov1, and D. I. Sokolov1 Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 2, pp. 129-138, June, 2020 Original article submitted September 6, 2019 Morphological properties and the size of microvesicles were assessed using atomic force microscopy, electron microscopy, and granulometric analysis. As these methods require significant numbers of microvesicles, we chose microvesicles derived from cell lines for our research. Key Words: microvesicles; transmission electron microscopy; atomic force microscopy; granulometric analysis; cell lines Analysis of small biological objects, e.g. microvesicles, is often associated with not only their size, but also their heterogeneity and resolving power of the equipment. Microvesicles (MV) are a type of extracellular vesicle [22], subcellular objects encapsulated in a bilayer membrane and varying in size from 100 to 1000 nm [13]. These objects are produced by various eukaryotic cells [27] and are present in almost all human biological fluids: blood plasma, urine, pleural and synovial fluid, saliva, breast milk, ejaculate, and nasal lavage under normal and pathological conditions [22,24]. MV contain various surface and intracellular molecules (proteins, lipids, glycolipids, glycoproteins, and nucleic acids, including DNA, mRNA, and non-coding RNA), due to which they can transmit information to adjacent cells and tissues. It is believed that MV can act as a subcellular vector delivering signals to surrounding cells, thereby serving as an element of cell—cell communication [22]. It was shown that MV regulate various processes in the body, including coagulation, angiogenesis [25], and antigen presentation. D. O. Ott Research Institute of Obstetrics, Gynecology, and Reproductology; 2Resource Centre for the Molecular and Cell Technologies Development, 3Resource Centre for Optical and Laser Materials Research, 4Resource Centre for Microscopy and Microanalysis, St. Petersburg State University, St. Petersburg, Russia. Address for correspondence: [email protected]. K. L. Markova 1
They can also transfer chemokine receptors, cytokine receptors, and arachidonic acid to the recipient cells. MV can exhibit both anti-inflammatory [27] (induction of apoptosis and production of anti-inflammatory mediators by immune cells [8]) and proinflammatory activities (increase cell susceptibility to new stimuli). MV mediate extracellular transfer of receptors and their incorporation into the plasma membrane of recipient cells [24], activate the complement system, and promote leukocyte rolling and mobilization into the pathological focus [19]. Thus, MV participate in the pathogenesis of various diseases and inflammatory processes [3,8,21
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