Abstract
Extracellular vesicles (EVs) have received immense attention in the past decade for their diverse use in diagnosis and therapeutics. Enhancing our understanding of EVs and increasing the reliability and reproducibility of EV research demands the use of standard isolation procedures and multiple characterization methods. Here we describe the most commonly used EV isolation method involving ultracentrifugation, and various characterization methods that include nanoparticle tracking analysis, atomic force microscopy and electron microscopy, which measure the size, concentration, and morphology of EVs.
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References
Pan BT, Johnstone RM (1983) Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro: selective externalization of the receptor. Cell 33(3):967–978
Marcilla A, Trelis M, Cortes A, Sotillo J, Cantalapiedra F, Minguez MT, Valero ML, Sanchez del Pino MM, Munoz-Antoli C, Toledo R, Bernal D (2012) Extracellular vesicles from parasitic helminths contain specific excretory/secretory proteins and are internalized in intestinal host cells. PLoS One 7(9):e45974. https://doi.org/10.1371/journal.pone.0045974
Luga V, Zhang L, Viloria-Petit AM, Ogunjimi AA, Inanlou MR, Chiu E, Buchanan M, Hosein AN, Basik M, Wrana JL (2012) Exosomes mediate stromal mobilization of autocrine Wnt-PCP signaling in breast cancer cell migration. Cell 151(7):1542–1556. https://doi.org/10.1016/j.cell.2012.11.024
Bjorge IM, Kim SY, Mano JF, Kalionis B, Chrzanowski W (2017) Extracellular vesicles, exosomes and shedding vesicles in regenerative medicine—a new paradigm for tissue repair. Biomater Sci 6(1):60–78. https://doi.org/10.1039/c7bm00479f
Su SA, Xie Y, Fu Z, Wang Y, Wang JA, Xiang M (2017) Emerging role of exosome-mediated intercellular communication in vascular remodeling. Oncotarget 8(15):25700–25712. https://doi.org/10.18632/oncotarget.14878
Bui TM, Mascarenhas LA, Sumagin R (2018) Extracellular vesicles regulate immune responses and cellular function in intestinal inflammation and repair. Tissue Barriers 6(2):e1431038. https://doi.org/10.1080/21688370.2018.1431038
Lane RE, Korbie D, Hill MM, Trau M (2018) Extracellular vesicles as circulating cancer biomarkers: opportunities and challenges. Clin Transl Med 7(1):14. https://doi.org/10.1186/s40169-018-0192-7
Urabe F, Kosaka N, Kimura T, Egawa S, Ochiya T (2018) Extracellular vesicles: toward a clinical application in urological cancer treatment. Int J Urol 25(6):533–543. https://doi.org/10.1111/iju.13594
Xu R, Rai A, Chen M, Suwakulsiri W, Greening DW, Simpson RJ (2018) Extracellular vesicles in cancer—implications for future improvements in cancer care. Nat Rev Clin Oncol 15(10):617–638. https://doi.org/10.1038/s41571-018-0036-9
Keerthikumar S, Chisanga D, Ariyaratne D, Al Saffar H, Anand S, Zhao K, Samuel M, Pathan M, Jois M, Chilamkurti N, Gangoda L, Mathivanan S (2016) ExoCarta: a web-based compendium of Exosomal cargo. J Mol Biol 428(4):688–692. https://doi.org/10.1016/j.jmb.2015.09.019
Gardiner C, Di Vizio D, Sahoo S, Thery C, Witwer KW, Wauben M, Hill AF (2016) Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey. J Extracell Vesicles 5:32945. https://doi.org/10.3402/jev.v5.32945
Garcia-Manrique P, Matos M, Gutierrez G, Pazos C, Blanco-Lopez MC (2018) Therapeutic biomaterials based on extracellular vesicles: classification of bio-engineering and mimetic preparation routes. J Extracell Vesicles 7(1):1422676. https://doi.org/10.1080/20013078.2017.1422676
Tian Y, Ma L, Gong M, Su G, Zhu S, Zhang W, Wang S, Li Z, Chen C, Li L, Wu L, Yan X (2018) Protein profiling and sizing of extracellular vesicles from colorectal cancer patients via flow cytometry. ACS Nano 12(1):671–680. https://doi.org/10.1021/acsnano.7b07782
Dipesh K, Bokai Z, Iqbal R, Curtis M, Quan L, Wojciech C (2018) Probing chemical and mechanical nanodomains in copolymer nanorods with correlative atomic force microscopy—nano-correscopy. Part Part Syst Charact 35(6):1700409. https://doi.org/10.1002/ppsc.201700409
Szatanek R, Baj-Krzyworzeka M, Zimoch J, Lekka M, Siedlar M, Baran J (2017) The methods of choice for extracellular vesicles (EVs) characterization. Int J Mol Sci 18(6). https://doi.org/10.3390/ijms18061153
Kim SY, Khanal D, Tharkar P, Kalionis B, Chrzanowski W (2018) None of us is the same as all of us: resolving the heterogeneity of extracellular vesicles using single-vesicle, nanoscale characterization with resonance enhanced atomic force microscope infrared spectroscopy (AFM-IR). Nanoscale Horizons 3(4):430–438. https://doi.org/10.1039/C8NH00048D
Acknowledgments
The authors acknowledge The University of Sydney for the SOAR Fellowship for W.Ch. The authors acknowledge the facilities and the scientific and technical assistance of the Bosch Molecular Biology Facility and Australian Microscopy & Microanalysis Research Facility at the Australian Centre for Microscopy & Microanalysis, The University of Sydney.
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Kim, S.Y., Phan, T.H., Limantoro, C., Kalionis, B., Chrzanowski, W. (2019). Isolation and Characterization of Extracellular Vesicles from Mesenchymal Stromal Cells. In: Joglekar, M., Hardikar, A. (eds) Progenitor Cells. Methods in Molecular Biology, vol 2029. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9631-5_2
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