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Article

Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation

1
Department of Medical Physics, Marian Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, 30-348 Krakow, Poland
2
Malopolska Centre of Biotechnology, Jagiellonian University, 30-837 Krakow, Poland
3
Department of Glycoconjugate Biochemistry, Institute of Zoology and Biomedical Research, Jagiellonian University, 30-837 Krakow, Poland
4
Department of General Chemistry, Faculty of Chemistry, Jagiellonian University, 30-837 Krakow, Poland
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Department of Gynecological Endocrinology, Faculty of Medicine, Jagiellonian University Medical College, 31-007 Krakow, Poland
6
Department of Obstetrics and Perinatology, Faculty of Medicine, Jagiellonian University Medical College, 31-007 Krakow, Poland
*
Author to whom correspondence should be addressed.
Pharmaceutics 2020, 12(9), 872; https://doi.org/10.3390/pharmaceutics12090872
Received: 26 July 2020 / Revised: 29 August 2020 / Accepted: 6 September 2020 / Published: 13 September 2020
(This article belongs to the Special Issue Extracellular Vesicles as Drug Delivery Systems)
Recent years have brought great focus on the development of drug delivery systems based on extracellular vesicles (EVs). Considering the possible applications of EVs as drug carriers, the isolation process is a crucial step. To solve the problems involved in EV isolation, we developed and validated a new EV isolation method—low-vacuum filtration (LVF)—and compared it with two commonly applied procedures—differential centrifugation (DC) and ultracentrifugation (UC). EVs isolated from endothelial cell culture media were characterized by (a) Transmission Electron Microscopy (TEM), (b) Nanoparticle Tracking Analysis (NTA), (c) Western blot and (d) Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Additionally, the membrane surface was imaged with Environmental Scanning Electron Microscopy (ESEM). We found that LVF was a reproducible and efficient method for EV isolation from conditioned media. Additionally, we observed a correlation between ATR-FTIR spectra quality and EV and protein concentration. ESEM imaging confirmed that the actual pore diameter was close to the values calculated theoretically. LVF is an easy, fast and inexpensive EV isolation method that allows for the isolation of both ectosomes and exosomes from high-volume sources with good repeatability. We believe that it could be an efficient alternative to commonly applied methods. View Full-Text
Keywords: dialysis membrane; ectosomes; exosomes; FTIR; infrared spectroscopy; purification dialysis membrane; ectosomes; exosomes; FTIR; infrared spectroscopy; purification
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MDPI and ACS Style

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. https://doi.org/10.3390/pharmaceutics12090872

AMA Style

Drożdż A, Kamińska A, Surman M, Gonet-Surówka A, Jach R, Huras H, Przybyło M, Stępień EŁ. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics. 2020; 12(9):872. https://doi.org/10.3390/pharmaceutics12090872

Chicago/Turabian Style

Drożdż, Anna, Agnieszka Kamińska, Magdalena Surman, Agnieszka Gonet-Surówka, Robert Jach, Hubert Huras, Małgorzata Przybyło, and Ewa Łucja Stępień. 2020. "Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation" Pharmaceutics 12, no. 9: 872. https://doi.org/10.3390/pharmaceutics12090872

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