Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

doi:10.3390/pharmaceutics12090872

Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

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Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

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dc.contributor.author	Drożdż, Anna [USOS96343]	pl
dc.contributor.author	Kamińska, Agnieszka [SAP14013162]	pl
dc.contributor.author	Surman, Magdalena [USOS130920]	pl
dc.contributor.author	Gonet-Surówka, Agnieszka [SAP14030302]	pl
dc.contributor.author	Jach, Robert [SAP20001622]	pl
dc.contributor.author	Huras, Hubert [SAP20002413]	pl
dc.contributor.author	Przybyło, Małgorzata [SAP11116102]	pl
dc.contributor.author	Stępień, Ewa [SAP14015168]	pl
dc.date.accessioned	2020-09-25T08:59:42Z
dc.date.available	2020-09-25T08:59:42Z
dc.date.issued	2020	pl
dc.identifier.uri	https://ruj.uj.edu.pl/xmlui/handle/item/246531
dc.language	eng	pl
dc.rights	Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa	*
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/pl/legalcode	*
dc.title	Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation	pl
dc.type	JournalArticle	pl
dc.abstract.en	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.	pl
dc.subject.en	dialysis membrane	pl
dc.subject.en	ectosomes	pl
dc.subject.en	exosomes	pl
dc.subject.en	FTIR	pl
dc.subject.en	infrared spectroscopy	pl
dc.subject.en	purification	pl
dc.description.volume	12	pl
dc.description.number	9	pl
dc.identifier.doi	10.3390/pharmaceutics12090872	pl
dc.identifier.eissn	1999-4923	pl
dc.title.journal	Pharmaceutics	pl
dc.language.container	eng	pl
dc.affiliation	Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego	pl
dc.affiliation	Wydział Biologii : Instytut Zoologii i Badań Biomedycznych	pl
dc.affiliation	Wydział Lekarski : Klinika Położnictwa i Perinatologii	pl
dc.affiliation	Wydział Lekarski : Klinika Endokrynologii Ginekologicznej	pl
dc.affiliation	Wydział Chemii : Zakład Chemii Ogólnej	pl
dc.subtype	Article	pl
dc.identifier.articleid	872	pl
dc.rights.original	CC-BY; otwarte czasopismo; ostateczna wersja wydawcy; w momencie opublikowania; 0	pl
dc.identifier.project	ROD UJ / OP	pl
dc.cm.id	101077
.pointsMNiSW	[2020 A]: 100