Cargando…

Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes

Extracellular vesicles are nanoparticles secreted by cell and have been proposed as suitable markers to identify competent embryos produced in vitro. Characterizing EVs secreted by individual embryos is challenging because culture medium itself contributes to the pool of nanoparticles that are co-is...

Descripción completa

Detalles Bibliográficos
Autores principales: Melo-Báez, Bárbara, Mellisho, Edwin A., Cabezas, Joel, Velásquez, Alejandra E., Veraguas, Daniel, Escobar, Diego Andrés Caamaño, Castro, Fidel O., Rodríguez-Álvarez, Lleretny
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Colégio Brasileiro de Reprodução Animal 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189351/
https://www.ncbi.nlm.nih.gov/pubmed/34122648
http://dx.doi.org/10.1590/1984-3143-AR2020-0028
_version_ 1783705489867014144
author Melo-Báez, Bárbara
Mellisho, Edwin A.
Cabezas, Joel
Velásquez, Alejandra E.
Veraguas, Daniel
Escobar, Diego Andrés Caamaño
Castro, Fidel O.
Rodríguez-Álvarez, Lleretny
author_facet Melo-Báez, Bárbara
Mellisho, Edwin A.
Cabezas, Joel
Velásquez, Alejandra E.
Veraguas, Daniel
Escobar, Diego Andrés Caamaño
Castro, Fidel O.
Rodríguez-Álvarez, Lleretny
author_sort Melo-Báez, Bárbara
collection PubMed
description Extracellular vesicles are nanoparticles secreted by cell and have been proposed as suitable markers to identify competent embryos produced in vitro. Characterizing EVs secreted by individual embryos is challenging because culture medium itself contributes to the pool of nanoparticles that are co-isolated. To avoid this, culture medium must be depleted of nanoparticles that are present in natural protein source. The aim of this study was to evaluate if the culture medium subjected to nanoparticle depletion can support the proper in vitro development of bovine embryos. Zygotes were cultured in groups on depleted or control medium for 8 days. Nanoparticles from the medium were characterized by their morphology, size and expression of EVs surface markers. Isolated nanoparticles were labelled and added to depleted medium containing embryos at different developmental stages and evaluated after 24 hours at 2, 8-16 cells, morula and blastocyst stages. There were no statistical differences on blastocyst rate at day 7 and 8, total cell count neither blastocyst diameter between groups. However, morphological quality was better in blastocysts cultured in non-depleted medium and the expression of SOX2 was significantly lower whereas NANOG expression was significantly higher. Few nanoparticles from medium had a typical morphology of EVs but were positive to specific surface markers. Punctuated green fluorescence near the nuclei of embryonic cells was observed in embryos from all developmental stages. In summary, nanoparticles from culture medium are internalized by in vitro cultured bovine embryos and their depletion affects the capacity of medium to support the proper embryo development.
format Online
Article
Text
id pubmed-8189351
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Colégio Brasileiro de Reprodução Animal
record_format MEDLINE/PubMed
spelling pubmed-81893512021-06-11 Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes Melo-Báez, Bárbara Mellisho, Edwin A. Cabezas, Joel Velásquez, Alejandra E. Veraguas, Daniel Escobar, Diego Andrés Caamaño Castro, Fidel O. Rodríguez-Álvarez, Lleretny Anim Reprod Original Article Extracellular vesicles are nanoparticles secreted by cell and have been proposed as suitable markers to identify competent embryos produced in vitro. Characterizing EVs secreted by individual embryos is challenging because culture medium itself contributes to the pool of nanoparticles that are co-isolated. To avoid this, culture medium must be depleted of nanoparticles that are present in natural protein source. The aim of this study was to evaluate if the culture medium subjected to nanoparticle depletion can support the proper in vitro development of bovine embryos. Zygotes were cultured in groups on depleted or control medium for 8 days. Nanoparticles from the medium were characterized by their morphology, size and expression of EVs surface markers. Isolated nanoparticles were labelled and added to depleted medium containing embryos at different developmental stages and evaluated after 24 hours at 2, 8-16 cells, morula and blastocyst stages. There were no statistical differences on blastocyst rate at day 7 and 8, total cell count neither blastocyst diameter between groups. However, morphological quality was better in blastocysts cultured in non-depleted medium and the expression of SOX2 was significantly lower whereas NANOG expression was significantly higher. Few nanoparticles from medium had a typical morphology of EVs but were positive to specific surface markers. Punctuated green fluorescence near the nuclei of embryonic cells was observed in embryos from all developmental stages. In summary, nanoparticles from culture medium are internalized by in vitro cultured bovine embryos and their depletion affects the capacity of medium to support the proper embryo development. Colégio Brasileiro de Reprodução Animal 2021-05-10 /pmc/articles/PMC8189351/ /pubmed/34122648 http://dx.doi.org/10.1590/1984-3143-AR2020-0028 Text en Copyright © The Author(s). https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Melo-Báez, Bárbara
Mellisho, Edwin A.
Cabezas, Joel
Velásquez, Alejandra E.
Veraguas, Daniel
Escobar, Diego Andrés Caamaño
Castro, Fidel O.
Rodríguez-Álvarez, Lleretny
Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title_full Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title_fullStr Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title_full_unstemmed Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title_short Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
title_sort nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189351/
https://www.ncbi.nlm.nih.gov/pubmed/34122648
http://dx.doi.org/10.1590/1984-3143-AR2020-0028
work_keys_str_mv AT melobaezbarbara nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT mellishoedwina nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT cabezasjoel nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT velasquezalejandrae nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT veraguasdaniel nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT escobardiegoandrescaamano nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT castrofidelo nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes
AT rodriguezalvarezlleretny nanoparticlesfromculturemediaareinternalizedbyinvitroproducedbovineembryosanditsdepletionaffectexpressionofpluripotencygenes