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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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Isolation-of-Extracellular-Vesicles-from-Human-Follicular-Fluid-SizeExclusion-Chromatography-versus-UltracentrifugationBiomolecules.pdf | 1.75 MB | Adobe PDF | View/Open |
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