PMID- 30868953 OWN - NLM STAT- MEDLINE DCOM- 20200723 LR - 20200723 IS - 1875-5666 (Electronic) IS - 1566-5240 (Linking) VI - 19 IP - 4 DP - 2019 TI - Blood Circulating Exosomes Contain Distinguishable Fractions of Free and Cell-Surface-Associated Vesicles. PG - 273-285 LID - 10.2174/1566524019666190314120532 [doi] AB - BACKGROUND: Considering exosomes as intercellular transporters, inevitably interacting with the plasma membrane and the large available surface of blood cells, we wonder if a fraction of circulating exosomes is associated with the surface of blood cells. OBJECTIVE: The aim of this study was to develop an efficient protocol for isolating exosomes associated with the surface of blood cells and to further investigate the characteristics of this fraction in a healthy state and during the development of breast cancer, as well as its possible implication for use in diagnostic applications. METHODS: Blood samples were collected from Healthy Females (HFs) and breast cancer patients (BCPs). Exosomes extracted from blood plasma and eluted from the surface of blood cells were isolated by ultrafiltration with subsequent ultracentrifugation. RESULTS: Transmission Electron Microscopy (TEM), along with immunogold labeling, demonstrated the presence of exosomes among membrane-wrapped extracellular vesicles (EVs) isolated from both plasma and blood cell eluates. TEM, nanoparticle tracking analysis, and NanoOrange protein quantitation data showed that cell-associated exosomes constituted no less than 2/3 of total blood exosome number. Exosomes, ranging from 50-70 nm in size, prevailed in the blood of breast cancer patients, whereas smaller exosomes (30-50 nm) were mostly observed in the blood of healthy women. Analysis of specific proteins and RNAs in exosomes circulating in blood demonstrated the significant differences in the packing density of the polymers in exosomes of HFs and BCPs. Preliminary data indicated that detection of cancer-specific miRNA (miR-103, miR-191, miR-195) in exosomes associated with the fraction of red blood cells allowed to discriminate HFs and BCPs more precisely compared to cell-free exosomes circulating in plasma. CONCLUSION: Our data provide the basis for using blood cell-associated exosomes for diagnostic applications. CI - Copyright(c) Bentham Science Publishers; For any queries, please email at epub@benthamscience.net. FAU - Tamkovich, Svetlana AU - Tamkovich S AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. AD - Institute for Regenerative Medicine, Lomonosov Moscow State University, Moscow, Russian Federation. AD - Faculty of Natural Science, Novosibirsk State University, Novosibirsk, Russian Federation. FAU - Tutanov, Oleg AU - Tutanov O AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. FAU - Efimenko, Anastasia AU - Efimenko A AD - Institute for Regenerative Medicine, Lomonosov Moscow State University, Moscow, Russian Federation. FAU - Grigor'eva, Alina AU - Grigor'eva A AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. FAU - Ryabchikova, Elena AU - Ryabchikova E AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. AD - Faculty of Natural Science, Novosibirsk State University, Novosibirsk, Russian Federation. FAU - Kirushina, Natalia AU - Kirushina N AD - Department of Mammology, National Novosibirsk Regional Oncologic Dispensary, Novosibirsk, Russian Federation. FAU - Vlassov, Valentin AU - Vlassov V AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. AD - Faculty of Natural Science, Novosibirsk State University, Novosibirsk, Russian Federation. FAU - Tkachuk, Vsevolod AU - Tkachuk V AD - Institute for Regenerative Medicine, Lomonosov Moscow State University, Moscow, Russian Federation. FAU - Laktionov, Pavel AU - Laktionov P AD - Laboratory of Molecular Medicine, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Novosibirsk, Russian Federation. AD - Novosibirsk Research Institute of Circulation Pathology of Academician E.N. Meshalkin, Novosibirsk, Russian Federation. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - Netherlands TA - Curr Mol Med JT - Current molecular medicine JID - 101093076 RN - 0 (Biomarkers) RN - 0 (Circulating MicroRNA) RN - 0 (MicroRNAs) SB - IM MH - Aged MH - Biomarkers MH - Blood Cells/metabolism MH - Breast Neoplasms/genetics/metabolism/pathology MH - Cell Fractionation MH - Cell-Derived Microparticles/*metabolism/ultrastructure MH - Circulating MicroRNA MH - Exosomes/*metabolism/ultrastructure MH - Extracellular Vesicles/metabolism MH - Female MH - Humans MH - Liquid Biopsy MH - MicroRNAs/genetics/metabolism MH - Middle Aged MH - Neoplasm Staging MH - Secretory Vesicles/*metabolism/ultrastructure OTO - NOTNLM OT - Exosomes OT - blood cells OT - breast cancer OT - cell-associated exosomes OT - extracellular vesicles OT - miRNA. EDAT- 2019/03/15 06:00 MHDA- 2020/07/24 06:00 CRDT- 2019/03/15 06:00 PHST- 2018/12/03 00:00 [received] PHST- 2019/03/08 00:00 [revised] PHST- 2019/03/08 00:00 [accepted] PHST- 2019/03/15 06:00 [pubmed] PHST- 2020/07/24 06:00 [medline] PHST- 2019/03/15 06:00 [entrez] AID - CMM-EPUB-97323 [pii] AID - 10.2174/1566524019666190314120532 [doi] PST - ppublish SO - Curr Mol Med. 2019;19(4):273-285. doi: 10.2174/1566524019666190314120532.