Isolation and characterization of exosomes derived from buffalo oviductal epithelial cells
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Keywords:
Exosomes, In vitro culture, Oviductal epithelial cells (OECS)Abstract
In the present study, exosomes were successfully isolated from the conditioned medium of buffalo OECs cultured in vitro. Characterisation by nanoparticle analysis revealed that the size of the oviductal exosomes ranged between 40-150 nm. Imaging by Transmission electron microscopy showed that these exosomes exhibited circular/spherical morphology. The identity of isolated exosomes was further confirmed by analysing the expression of surface expressed tetraspannin markers (CD9, CD63, and CD81) by flow cytometry, and were found to be enriched with the tetraspannins analysed. As continuation of this work, further research to supplement the isolated oviductal exosomes to recreate the preimplantation physiological environment of the oviduct should be undertaken, thus improving in vitro culture conditions for buffalo embryos.
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References
Alminana C and Bauersachs S. 2019. Extracellular vesicles in the oviduct: progress, challenges and implications for the reproductive success. Bioengineering 6(2): 32. DOI: https://doi.org/10.3390/bioengineering6020032
Capra E and Lange-Consiglio A. 2020. The biological function of extracellular vesicles during fertilization, early embryo- maternal crosstalk and their involvement in reproduction: Review and overview. Biomolecules 10(11): 1510. DOI: https://doi.org/10.3390/biom10111510
Cordova A, Perreau C, Uzbekova S, Ponsart C, Locatelli Y and Mermillod P. 2014. Development rate and gene expression of IVP bovine embryos cocultured with bovine oviduct epithelial cells at early or late stage of preimplantation development. Theriogenology 81(9): 1163–73. DOI: https://doi.org/10.1016/j.theriogenology.2014.01.012
García E V, Hamdi M, Barrera A D, Sanchez-Calabuig M J, Gutierrez-Adan A and Rizos D. 2017. Bovine embryo-oviduct interaction in vitro reveals an early cross-talk mediated by BMP signalling. Reproduction 153(5): 631–43. DOI: https://doi.org/10.1530/REP-16-0654
Hasan M M, Reshi Q U A, Lättekivi F, Viil J, Godakumara K, Dissanayake K, Andronowska A, Jaakma U and Fazeli A. 2021. Bovine follicular fluid derived extracellular vesicles modulate the viability, capacitation, and acrosome reaction of bull spermatozoa. Biology 10(11): 1154. DOI: https://doi.org/10.3390/biology10111154
Kowal J, Arras G, Colombo M, Jouve M, Morath J P, Primdal- Bengtson B, Dingli F, Loew D, Tkach M and Thery C. 2016.
Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of National Academy of Science, USA 113: 968–77.
Lopera-Vasquez R, Hamdi M, Fernandez- Fuertes B, Maillo V, Beltran-Brena P and Calle A, Redruello A, Lopez-Martin S, Gutierrez-Adan A, Yanez-Mo M, Ramirez M A and Rizos D. 2016. Extracellular vesicles from BOEC in in vitro embryo development and quality. PLoS ONE 11(2): e0148083. DOI: https://doi.org/10.1371/journal.pone.0148083
Matsuno Y, Onuma A, Fujioka Y A, Yasuhara K, Fujii W, Naito K and Sugiura K. 2017. Effects of exosome-like vesicles on cumulus expansion in pigs in vitro. Journal of Reproduction and Development 63(1): 51–58. DOI: https://doi.org/10.1262/jrd.2016-124
Mellisho E A, Velasquez A E, Nuñez M J, Cabezas J G, Cueto J A, Fader C, Castro F O and Rodriguez-Alvarez L. 2017. Identification and characteristics of extracellular vesicles from bovine blastocysts produced in vitro. PLoS ONE 12(5): e0178306. DOI: https://doi.org/10.1371/journal.pone.0178306
Melo-Baez B, Mellisho E A, Cabezas J, Velasquez AE, Veraguas D, Escobar D A C, Castro F O and Rodriguez-Alvarez L. 2020. Nanoparticles from culture media are internalized by in vitro- produced bovine embryos and its depletion affect expression of pluripotency genes. Animal Reproduction 18(1): e20200028. DOI: https://doi.org/10.1590/1984-3143-ar2020-0028
Mondejar I, Aviles M and Coy P. 2013. The human is an exception to the evolutionarily-conserved phenomenon of pre- fertilization zona pellucida resistance to proteolysis induced by oviductal fluid. Human Reproduction 28(3): 718–28. DOI: https://doi.org/10.1093/humrep/des423
Ni Z, Zhou S, Li S, Kuang L, Chen H and Luo X, Ouyang J, He M, Du X and Chen L. 2020. Exosomes: Roles and therapeutic potential in osteoarthritis. Bone Research 8(1): 25. Pavani K C, Alminana C, Wydooghe E, Catteeuw M, Ramirez M A, Mermillod P D, Rizos D and Van Soom A. 2017. Emerging role of extracellular vesicles in communication of preimplantation embryos in vitro. Reproduction, Fertility and Development 29(1): 66–83. DOI: https://doi.org/10.1071/RD16318
Szatanek R, Baj-Krzyworzeka M, Zimoch J, Lekka M, Siedlar M and Baran J. 2017. The Methods of choice for extracellular vesicles (EVs) characterization. International Journal of Molecular Science 18(6): 1153. DOI: https://doi.org/10.3390/ijms18061153
Thery C, Witwer K W, Aikawa E, Alcaraz M J, Anderson J D, Andriantsitohaina R, Antoniou A, Arab T, Archer Fatkin-Smith G K, Ayre D C, Bach J M, Bachurski D, Baharvand H, Balaj L, Baldacchino S, Bauer N N, Baxter A A, Bebawy M, Beckham C, Zavec A B, et al. 2018. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. Journal of Extracellular Vesicles 7(1): 1535750. DOI: https://doi.org/10.1080/20013078.2018.1535750
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