Please use this identifier to cite or link to this item: https://hdl.handle.net/10316/113957
Title: Isolation of Extracellular Vesicles from Human Follicular Fluid: Size-Exclusion Chromatography versus Ultracentrifugation
Authors: Soares, Maria 
Pinto, Maria M. 
Nobre, Rui J. 
Almeida, Luís Pereira de 
Rasteiro, Maria da Graça 
Santos, Teresa Almeida 
Santos, João Ramalho 
Sousa, Ana Paula 
Keywords: follicular fluid; extracellular vesicles; ultracentrifugation; size-exclusion chromatography
Issue Date: 2-Feb-2023
Publisher: MDPI
Project: CENTRO-01-0145-FEDER-028871 
CENTRO-01-0145-FEDER-000012 
POCI-01-0145-FEDER-007440 
2020.06180.BD 
2022.11089.BD 
UIDB/00102/2020 
metadata.degois.publication.title: Biomolecules
metadata.degois.publication.volume: 13
metadata.degois.publication.issue: 2
Abstract: Follicular fluid (FF) is the microenvironment where a growing oocyte develops. Intrafollicular communication ensures oocyte competence and is carried out through paracrine signaling, the exchange of molecules via gap junctions, and the trafficking of extracellular vesicles (EVs). The study of FF-derived EVs is important for both translational and fundamental research in the female reproductive field. This study aimed to compare the efficacy and purity of two EV isolation methods: size-exclusion chromatography (SEC) and ultracentrifugation (UC). EVs isolated using SEC and UC were compared regarding their size and concentration using dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA); protein contamination was assessed with microBCA; specific EV markers were detected with Western blot, and EV morphology was studied with transmission electron microscopy (TEM). Our results show that although both techniques isolated small EVs, a significantly increased yield in particle number was clear with UC compared with SEC. On the other hand, SEC generated purer EVs with fewer protein contaminants and aggregates. In conclusion, the selection of the most suited approach to isolate EVs must be conducted considering the degree of recovery, purity, and downstream application of the isolated EVs.
URI: https://hdl.handle.net/10316/113957
ISSN: 2218-273X
DOI: 10.3390/biom13020278
Rights: openAccess
Appears in Collections:FCTUC Eng.Química - Artigos em Revistas Internacionais
I&D CERES - Artigos em Revistas Internacionais
I&D CNC - Artigos em Revistas Internacionais
IIIUC - Artigos em Revistas Internacionais
FFUC- Artigos em Revistas Internacionais

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