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. 2022 Nov 1;44(11):5363-5378.
doi: 10.3390/cimb44110363.

Comparative Analysis of Natural and Cytochalasin B-Induced Membrane Vesicles from Tumor Cells and Mesenchymal Stem Cells

Zarema Gilazieva  1 , Daria Chulpanova  1 , Aleksei Ponomarev  1 , Ivan Filin  1 , Ekaterina Garanina  1 , Albert Rizvanov  1 , Valeriya Solovyeva  1
Affiliations

Comparative Analysis of Natural and Cytochalasin B-Induced Membrane Vesicles from Tumor Cells and Mesenchymal Stem Cells

Zarema Gilazieva et al. Curr Issues Mol Biol. .

Abstract

To date, there are numerous protocols for the isolation of extracellular vesicles (EVs). Depending on the isolation method, it is possible to obtain vesicles with different characteristics, enriched with specific groups of proteins, DNA and RNA, which affect similar types of cells in the opposite way. Therefore, it is important to study and compare methods of vesicle isolation. Moreover, the differences between the EVs derived from tumor and mesenchymal stem cells are still poorly understood. This article compares EVs from human glioblastoma cells and mesenchymal stem cells (MSCs) obtained by two different methods, ultracentrifugation and cytochalasin B-mediated induction. The size of the vesicles, the presence of the main EV markers, the presence of nuclear and mitochondrial components, and the molecular composition of the vesicles were determined. It has been shown that EVs obtained by both ultracentrifugation and cytochalasin B treatment have similar features, contain particles of endogenous and membrane origin and can interact with monolayer cultures of tumor cells.

Keywords: cytochalasin B; extracellular vesicles; glioblastoma; mesenchymal stem cells; tumor microenvironment; ultracentrifugation.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; and in the decision to publish the results.

Figures

Figure 1
Figure 1
Production of cytochalasin B-induced membrane vesicles and natural extracellular vesicles. Isolation procedure design of CIMVs and NEVs.
Figure 2
Figure 2
Characteristics of MSCs. (A) Directed differentiation of isolated MSCs into adipogenic, chondrogenic and osteogenic directions. Scale: 100 µm. (B) Immunophenotypic characteristics of MSCs isolated from human adipose tissue. The graphs represent the percentage of positive cells. Negative control−a mixture of hematopoietic cell markers (CD11b, CD19, CD34, CD45, HLA-DR and CD45). Data are shown as the mean ± SD (n = 6) of two biological replicates.
Figure 3
Figure 3
Morphology and size of CIMVs and NEVs isolated from MCSs and SNB-19. The data were obtained using scanning electron microscopy, scale: 1 µm, magnification: ×10.00 k. Data are shown as the mean ± SD (n = 10) of two biological replicates.
Figure 4
Figure 4
Analysis of the expression of typical EV markers on the CIMVs and NEVs. Data are shown as the mean percentage ± SD (n = 6) of two biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Figure 5
Figure 5
Heatmap of cytokine/chemokine concentrations in the cell and extracellular vesicle lysate. (A) Samples of MSCs. (B) Samples of SNB-19. All the samples were normalized relative to the total protein concentration. Data are shown as the mean ± SD (n = 6) of two biological replicates.
Figure 6
Figure 6
(A) Analysis of the presence of the mitochondrial component in CIMVs and NEVs. (B) Analysis of the presence of a nuclear component in CIMVs and NEVs. (C) Percentage of CIMVs and NEVs stained with membrane dye. Data are shown as the mean percentage ± SD (n = 6) of two biological replicates. * p < 0.05, **** p < 0.0001.
Figure 7
Figure 7
Confocal microscopy of HCT-15 and MCF7 tumor cells in monolayer culture. Cell nuclei were stained with DAPI (blue). CIMVs and NEVs stained with DiO membrane stain (green) are shown by arrows. Scale: 100 µm.
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References

    1. Gilazieva Z., Ponomarev A., Rizvanov A., Solovyeva V. The Dual Role of Mesenchymal Stromal Cells and Their Extracellular Vesicles in Carcinogenesis. Biology. 2022;11:813. doi: 10.3390/biology11060813. - DOI - PMC - PubMed
    1. Choi D.S., Kim D.K., Kim Y.K., Gho Y.S. Proteomics of extracellular vesicles: Exosomes and ectosomes. Mass Spectrom. Rev. 2015;34:474–490. doi: 10.1002/mas.21420. - DOI - PubMed
    1. Delcayre A., Estelles A., Sperinde J., Roulon T., Paz P., Aguilar B., Villanueva J., Khine S., Le Pecq J.B. Exosome Display technology: Applications to the development of new diagnostics and therapeutics. Blood Cells Mol. Dis. 2005;35:158–168. doi: 10.1016/j.bcmd.2005.07.003. - DOI - PubMed
    1. Simhadri V.R., Reiners K.S., Hansen H.P., Topolar D., Simhadri V.L., Nohroudi K., Kufer T.A., Engert A., Pogge von Strandmann E. Dendritic cells release HLA-B-associated transcript-3 positive exosomes to regulate natural killer function. PLoS ONE. 2008;3:e3377. doi: 10.1371/journal.pone.0003377. - DOI - PMC - PubMed
    1. Koniusz S., Andrzejewska A., Muraca M., Srivastava A.K., Janowski M., Lukomska B. Extracellular Vesicles in Physiology, Pathology, and Therapy of the Immune and Central Nervous System, with Focus on Extracellular Vesicles Derived from Mesenchymal Stem Cells as Therapeutic Tools. Front. Cell. Neurosci. 2016;10:109. doi: 10.3389/fncel.2016.00109. - DOI - PMC - PubMed

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