Diversity of Extracellular Vesicles in Human Follicular Fluid: Morphological Analysis and Quantification

被引:17
|
作者
Neyroud, Anne-Sophie [1 ,2 ]
Chiechio, Regina Maria [3 ,4 ]
Moulin, Gregory [1 ,3 ]
Ducarre, Solene [3 ]
Heichette, Claire [5 ]
Dupont, Aurelien [5 ]
Budzynski, Mathieu [1 ]
Even-Hernandez, Pascale [3 ]
Lo Faro, Maria Jose [4 ]
Yefimova, Marina [1 ,2 ]
Marchi, Valerie [3 ]
Ravel, Celia [1 ,2 ]
机构
[1] CHU Rennes, Serv Biol Reprod CECOS, F-35000 Rennes, France
[2] Inst Rech Sante, Environm & Travail IRSET UMR S 1085, F-35000 Rennes, France
[3] Univ Rennes 1, CNRS UMR 6226, Inst Sci Chim Rennes, Ave Gen Leclerc, F-35042 Rennes, France
[4] Univ Catania, Dipartimento Fis & Astron Ettore Majorana, Via Santa Sofia 64, I-95123 Catania, Italy
[5] Univ Rennes, UMS 3480, US S 018, BIOSIT,CNRS,INSERM, F-35000 Rennes, France
关键词
extracellular vesicles; follicular fluid; human; oocyte; morphology; EXOSOMES;
D O I
10.3390/ijms231911676
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The oocyte microenvironment constituted by the follicular fluid (FF) is a key for the optimal development of female gametes. Its composition reflects the physiological state of the ovarian follicle. The particularity of FF is to contain a huge diversity of extracellular vesicles specific to women, in the same way as seminal plasma in men. Here, we described and compared morphological aspects of broad subcategories of human FF-related Extracellular Vesicles (EVs). EVs participate in physiological and pathological processes and have potential applications in diagnostics or therapeutics. EVs isolated from FF are involved in different biological functions related to follicular growth, oocyte maturation, and embryo development. However, knowledge on the morphology of FF-derived EVs is limited, mainly due to their sub-micrometer size and to intrinsic limitations in methods applied for their characterization. The aim of this study was to provide a comprehensive morphological description of EVs from FF of healthy subjects and quantification. EVs separation was realized by centrifugation, with comparison of the EV yield obtained from differential centrifugation and one-step ultracentrifugation. Cryo-Transmission Electron Microscopy was used to reveal the morphology, size, and phenotype of EVs. Dynamic Light Scattering (DLS) and Nanoparticle Tracking Analysis (NTA) were used to quantify and analyze the size distribution for each centrifugation step. We performed a comprehensive inventory of human follicular fluid EVs. We show that human FF contains a huge diversity of EVs. This study brings novel insights on EVs from normal FF and provides a reference for further studies of EVs in ovarian diseases.
引用
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页数:13
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