Please use this identifier to cite or link to this item:
https://elib.bsu.by/handle/123456789/289034
Title: | Cm-dil staining and sec of plasma as an approach to increase sensitivity of extracellular nanovesicles quantification by bead-assisted flow cytometry |
Authors: | Nikiforova, Nadezhda Chumachenko, Maria Nazarova, Inga Zabegina, Lidia Slyusarenko, Maria Sidina, Elena Malek, Anastasia |
Keywords: | ЭБ БГУ::ЕСТЕСТВЕННЫЕ И ТОЧНЫЕ НАУКИ::Биология |
Issue Date: | 2021 |
Publisher: | MDPI AG |
Citation: | Membr 2021;11(7). |
Abstract: | The quantification of the specific disease-associated populations of circulating extracellular membrane nanovesicles (ENVs) has opened up new opportunities for liquid biopsy in cancer and other chronic diseases. However, the sensitivity of such methods is mediated by an optimal combination of the isolation and labeling approaches, and is not yet sufficient for routine clinical application. The presented study aimed to develop, characterize, and explore a new approach to non-specific ENV staining, followed by size-exclusive chromatography (SEC), which allows us to increase the sensitivity of bead-assisted flow cytometry. Plasma from healthy donors was purified from large components, stained with lipophilic CM-Dil dye, and fractionated by means of SEC. The obtained fractions were analyzed in terms of particle size and concentration using NTA, as well as vesicular markers and plasma protein content via dot-blotting. We characterized the process of CM-Dil-stained plasma fractionation in detail and indicated the fractions with optimal characteristics. Finally, we explored the sensitivity of on-bead flow cytometry for the analysis of specific populations of plasma ENVs and demonstrated the advantages and limitations of the proposed technique. |
URI: | https://elib.bsu.by/handle/123456789/289034 |
DOI: | 10.3390/membranes11070526 |
Scopus: | 85111683253 |
Sponsorship: | This research was funded by the Ministry of Health of the Russian Federation in the framework of the “Development and clinical testing of methods for isolation, analysis and modification of the circulating plasma nanovesicles with the aim of personalized selection and increasing the effectiveness of standard regimens of systemic therapy for oncological diseases” project № 056-00108-21-02/121032300202-6, 2021–2023. |
Licence: | info:eu-repo/semantics/openAccess |
Appears in Collections: | Статьи биологического факультета |
Files in This Item:
File | Description | Size | Format | |
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membranes-11-00526.pdf | 2,24 MB | Adobe PDF | View/Open |
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