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dc.contributor.author
Mellisho, Edwin A.
dc.contributor.author
Velásquez, Alejandra E.
dc.contributor.author
Nuñez, María J.
dc.contributor.author
Cabezas, Joel G.
dc.contributor.author
Cueto, Juan Agustin
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dc.contributor.author
Fader Kaiser, Claudio Marcelo
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dc.contributor.author
Castro, Fidel O.
dc.contributor.author
Rodriguez Alvarez, Lleretny
dc.date.available
2018-06-19T19:50:00Z
dc.date.issued
2017-05
dc.identifier.citation
Mellisho, Edwin A.; Velásquez, Alejandra E.; Nuñez, María J.; Cabezas, Joel G.; Cueto, Juan Agustin; et al.; Identification and characteristics of Extracellular vesicles from bovine blastocysts produced in vitro; Public Library of Science; Plos One; 12; 5; 5-2017; 1-15
dc.identifier.issn
1932-6203
dc.identifier.uri
http://hdl.handle.net/11336/49431
dc.description.abstract
Extracellular vesicles (EVs) have been identified within different body fluids and cell culture media. However, there is very little information on the secretion of these vesicles during early embryonic development. The aims of this work were first to demonstrate the secretion of extracellular vesicles by pre-implantation bovine embryos and second to identify and characterize the population of EVs secreted by bovine blastocysts during the period from day seven to nine of embryo culture and its correlation with further embryo development up to day 11. Bovine embryos were produced by in vitro fertilization (IVF) or parthenogenetic activation (PA) and cultured until blastocyst stage. Blastocyst selection was performed at day 7 post IVF/PA considering two variables: stage of development and quality of embryos. Selected blastocysts were cultured in vitro for 48 hours in groups (exp. 1) or individually (exp. 2) in SOF media depleted of exosomes. At day 9 post IVF/PA the media was collected and EVs isolated by ultracentrifugation. Transmission electron microscopy revealed the presence of heterogeneous vesicles of different sizes and population: microvesicles (MVs) and exosomes (EXs) of rounded shape, enclosed by a lipid bi-layer and ranging from 30 to 385 nm of diameter. Flow cytometry analysis allowed identifying CD63 and CD9 proteins as exosome markers. Nanoparticle tracking analysis generated a large number of variables, which required the use of multivariate statistics. The results indicated that the concentration of vesicles is higher in those blastocysts with arrested development from day 9 up to day 11 of in vitro development (6.7 x 108 particles/ml) derived from IVF (p <0.05), compared to PA blastocysts (4.7 x 108 particles/ml). Likewise, the profile (concentration and diameter) of particles secreted by embryos derived from IVF were different from those secreted by PA embryos. In conclusion, we demonstrated that bovine blastocysts secrete MVs/EXs to the culture media. Data suggest that characteristics of the population of EVs vary depending on embryo competence.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Public Library of Science
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Exosomes
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Mvbs
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Electron Microscopy
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Otras Ciencias Biológicas
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Identification and characteristics of Extracellular vesicles from bovine blastocysts produced in vitro
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-06-13T16:54:36Z
dc.journal.volume
12
dc.journal.number
5
dc.journal.pagination
1-15
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
San Francisco
dc.description.fil
Fil: Mellisho, Edwin A.. Universidad de Concepción; Chile
dc.description.fil
Fil: Velásquez, Alejandra E.. Universidad de Concepción; Chile
dc.description.fil
Fil: Nuñez, María J.. Universidad de Concepción; Chile
dc.description.fil
Fil: Cabezas, Joel G.. Universidad de Concepción; Chile
dc.description.fil
Fil: Cueto, Juan Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
dc.description.fil
Fil: Fader Kaiser, Claudio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
dc.description.fil
Fil: Castro, Fidel O.. Universidad de Concepción; Chile
dc.description.fil
Fil: Rodriguez Alvarez, Lleretny. Universidad de Concepción; Chile
dc.journal.title
Plos One
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://doi.org/10.1371/journal.pone.0178306
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0178306
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