Isolation and Characterization of Functionally Active Extracellular Vesicles from Culture Medium Conditioned by Bovine Embryos In Vitro

被引：57
作者：
Pavani, Krishna Chaitanya [1 ]
Hendrix, An [2 ,3 ]
Van Den Broeck, Wim [4 ]
Couck, Liesbeth [4 ]
Szymanska, Katarzyna [5 ]
Lin, Xiaoyuan [6 ]
De Koster, Jenne [1 ]
Van Soom, Ann [1 ]
Leemans, Bart [1 ]
机构：
[1] Univ Ghent, Fac Vet Med, Dept Reprod Obstet & Herd Hlth, Salisburylaan 133, B-9820 Merelbeke, Belgium
[2] Univ Ghent, Dept Radiat Oncol & Expt Canc Res, Lab Expt Canc Res, C Heymanslaan 10, B-9000 Ghent, Belgium
[3] CRIG, C Heymanslaan 10, B-9000 Ghent, Belgium
[4] Univ Ghent, Fac Vet Med, Dept Morphol, Salisburylaan 133, B-9820 Merelbeke, Belgium
[5] Univ Ghent, Fac Med & Hlth Sci, Dept Basic Med Sci, Physiol Grp, Pintelaan 185, B-9000 Ghent, Belgium
[6] Univ Ghent, Fac Vet Med, Dept Nutr Genet & Ethol, B-9820 Merelbeke, Belgium
来源：
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2019年 / 20卷 / 01期
基金：
欧盟地平线“2020”;
关键词：
embryo-embryo communication; extracellular vesicles; embryo culture; bovine; Optiprep (TM) density gradient ultracentrifugation; uptake of embryo-secreted EVs; ZONA-PELLUCIDA; EXOSOMES; BLASTOCYSTS; MICROVESICLES; SUPPLEMENTS; COCULTURE;
D O I：
10.3390/ijms20010038
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：
Extracellular vesicles (EVs) play a possible role in cell-cell communication and are found in various body fluids and cell conditioned culture media. The aim of this study was to isolate and characterize EVs in culture medium conditioned by bovine embryos in group and to verify if these EVs are functionally active. Initially, ultracentrifuged bovine serum albumin (BSA) containing medium was selected as suitable EV-free embryo culture medium. Next, EVs were isolated from embryo conditioned culture medium by OptiPrep(TM) density gradient ultracentrifugation. Isolated EVs were characterized by nanoparticle tracking analysis, western blotting, transmission, and immunoelectron microscopy. Bovine embryo-derived EVs were sizing between 25-230 nm with an average concentration of 236.5 +/- 1.27 x 10(8) particles/mL. Moreover, PKH67 EV pre-labeling showed that embryo-secreted EVs were uptaken by zona-intact bovine embryos. Since BSA did not appear to be a contaminating EV source in culture medium, EV functionality was tested in BSA containing medium. Individual embryo culture in BSA medium enriched with EVs derived from conditioned embryo culture medium showed significantly higher blastocyst rates at day 7 and 8 together with a significantly lower apoptotic cell ratio. In conclusion, our study shows that EVs play an important role in inter embryo communication during bovine embryo culture in group.
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页数：22
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