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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Colégio Brasileiro de Reprodução Animal
2021
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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 |
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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 |
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