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Low Vacuum Filtration Method as an Alternative Extracellular Vesicle Concentration Method: Comparison with Ultracentrifugation and Differential Centrifugation
Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics2020, 12, 872.
Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics 2020, 12, 872.
Cite as:
Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics2020, 12, 872.
Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics 2020, 12, 872.
Abstract
Recent years brought great focus in the field of development of extracellular vesicles (EVs) based drug-delivery systems. Considering possible applications of EVs as a drug carriers the isolation process is a crucial step. To solve problems related with EV isolation, we created and validated a new EVs isolation method – Low Vacuum Filtration (LVF) and compared it with two commonly applied procedures - differential centrifugation (DC) and ultracentrifugation (UC). EVs isolated from endothelial cells culture media have been characterized by a) transmission electron microscopy (TEM) b) nanoparticle tracking analysis (NTA), c) western blot and d) Fourier-Transform Infrared Spectroscopy (FTIR). Additionally, the membrane surface have been imaged with Environmental Scanning Electron Microscopy (ESEM). We showed that LVF is reproducible and efficient method for EVs isolation form conditioned media. Additionally, we observed correlation between ATR-FTIR spectra quality and the EVs and proteins concentration. ESEM imaging confirmed that actual pore diameter are close to the values calculated theoretically. LVF method is an easy, fast and inexpensive EVs isolation method which allows for isolation of both ectosomes and exosomes from high volume sources with good repeatability. We think that it could be an efficient alternative for commonly applied methods.
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.