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

2020
journal article
article
dc.abstract.enRecent 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.affiliationWydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiegopl
dc.affiliationWydział Biologii : Instytut Zoologii i Badań Biomedycznychpl
dc.affiliationWydział Lekarski : Klinika Położnictwa i Perinatologiipl
dc.affiliationWydział Lekarski : Klinika Endokrynologii Ginekologicznejpl
dc.affiliationWydział Chemii : Zakład Chemii Ogólnejpl
dc.cm.id101077
dc.contributor.authorDrożdż, Anna - 164492 pl
dc.contributor.authorKamińska, Agnieszka - 241454 pl
dc.contributor.authorSurman, Magdalena - 192235 pl
dc.contributor.authorGonet-Surówka, Agnieszka - 162000 pl
dc.contributor.authorJach, Robert - 129777 pl
dc.contributor.authorHuras, Hubert - 129736 pl
dc.contributor.authorPrzybyło, Małgorzata - 131574 pl
dc.contributor.authorStępień, Ewa - 161583 pl
dc.date.accessioned2020-09-25T08:59:42Z
dc.date.available2020-09-25T08:59:42Z
dc.date.issued2020pl
dc.date.openaccess0
dc.description.accesstimew momencie opublikowania
dc.description.number9pl
dc.description.versionostateczna wersja wydawcy
dc.description.volume12pl
dc.identifier.articleid872pl
dc.identifier.doi10.3390/pharmaceutics12090872pl
dc.identifier.eissn1999-4923pl
dc.identifier.projectROD UJ / OPpl
dc.identifier.urihttps://ruj.uj.edu.pl/xmlui/handle/item/246531
dc.languageengpl
dc.language.containerengpl
dc.rightsUdzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa*
dc.rights.licenceCC-BY
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/legalcode.pl*
dc.share.typeotwarte czasopismo
dc.subject.endialysis membranepl
dc.subject.enectosomespl
dc.subject.enexosomespl
dc.subject.enFTIRpl
dc.subject.eninfrared spectroscopypl
dc.subject.enpurificationpl
dc.subtypeArticlepl
dc.titleLow-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugationpl
dc.title.journalPharmaceuticspl
dc.typeJournalArticlepl
dspace.entity.typePublication
dc.abstract.enpl
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.
dc.affiliationpl
Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego
dc.affiliationpl
Wydział Biologii : Instytut Zoologii i Badań Biomedycznych
dc.affiliationpl
Wydział Lekarski : Klinika Położnictwa i Perinatologii
dc.affiliationpl
Wydział Lekarski : Klinika Endokrynologii Ginekologicznej
dc.affiliationpl
Wydział Chemii : Zakład Chemii Ogólnej
dc.cm.id
101077
dc.contributor.authorpl
Drożdż, Anna - 164492
dc.contributor.authorpl
Kamińska, Agnieszka - 241454
dc.contributor.authorpl
Surman, Magdalena - 192235
dc.contributor.authorpl
Gonet-Surówka, Agnieszka - 162000
dc.contributor.authorpl
Jach, Robert - 129777
dc.contributor.authorpl
Huras, Hubert - 129736
dc.contributor.authorpl
Przybyło, Małgorzata - 131574
dc.contributor.authorpl
Stępień, Ewa - 161583
dc.date.accessioned
2020-09-25T08:59:42Z
dc.date.available
2020-09-25T08:59:42Z
dc.date.issuedpl
2020
dc.date.openaccess
0
dc.description.accesstime
w momencie opublikowania
dc.description.numberpl
9
dc.description.version
ostateczna wersja wydawcy
dc.description.volumepl
12
dc.identifier.articleidpl
872
dc.identifier.doipl
10.3390/pharmaceutics12090872
dc.identifier.eissnpl
1999-4923
dc.identifier.projectpl
ROD UJ / OP
dc.identifier.uri
https://ruj.uj.edu.pl/xmlui/handle/item/246531
dc.languagepl
eng
dc.language.containerpl
eng
dc.rights*
Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa
dc.rights.licence
CC-BY
dc.rights.uri*
http://creativecommons.org/licenses/by/4.0/legalcode.pl
dc.share.type
otwarte czasopismo
dc.subject.enpl
dialysis membrane
dc.subject.enpl
ectosomes
dc.subject.enpl
exosomes
dc.subject.enpl
FTIR
dc.subject.enpl
infrared spectroscopy
dc.subject.enpl
purification
dc.subtypepl
Article
dc.titlepl
Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation
dc.title.journalpl
Pharmaceutics
dc.typepl
JournalArticle
dspace.entity.type
Publication

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