Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification

Autores
Rios, Glenda Laura; Suqueli García, María Florencia; Manrique, Rodrigo J.; Buschiazzo, Jorgelina
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Long-term conservation of livestock female genetic resources has led to increased interest in oocyte cryopreservation. However, the challenges of preserving large-volume cells and the lack of standardization in vitrification media and cryo-devices for domestic species remain critical areas of ongoing research. This study aimed to assess the impact of in vitro maturation medium composition and vitrification devices on cell death and membrane fusion competence of vitrified bovine oocytes, comparing a chemically undefined versus a synthetic maturation medium supplemented with linoleic acid (LA), and using both tubular (open-pulled straw) and surface vitrification devices (Cryotech®). In vitro maturation of bovine oocytes under serum-free conditions improved post-vitrification survival, particularly by preserving membrane integrity. Additionally, oocytes matured in a synthetic medium containing epidermal growth factor and hyaluronic acid, and vitrified using a surface device, exhibited increased viability and reduced caspase activation. Supplementation of this medium with 43 μM LA did not compromise oocyte viability or apoptotic status. In contrast, higher concentrations (100 μM) induced significant apoptosis and disrupted membrane integrity, impairing tolerance to cryoprotectant agents. We also identified two distinct localization patterns of the transcription factor OCT4 in matured oocytes, with vitrification promoting the diffuse pattern. Supplementation with LA was insufficient to mitigate the effects of vitrification on OCT4 distribution. Unlike 43 μM, maturation with 100 μM LA increased the proportion of oocytes exhibiting the diffuse OCT4 pattern following exposure to cryoprotectant agents under non-vitrified conditions. Zona pellucida-free oocytes co-incubated with sperm demonstrated that 43 μM LA better preserved the original sperm adhesion and fusion pattern post-vitrification, although fertilization rate was not improved. These findings demonstrate that optimized maturation conditions—comprising serum-free media, membrane fluidity-modulating lipids (43 μM LA), and the use of surface cryo-devices—significantly enhance oocyte quality, post-warming survival, and fusion competence, all of which are critical for improving fertilization outcomes following vitrification.
EEA Balcarce
Fil: Ríos, Glenda Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Suqueli García, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Suqueli García, María Florencia. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina
Fil: Manrique, Rodrigo J. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Buschiazzo, Jorgelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fuente
Frontiers in Veterinary Science 12: 1-14 (03 July 2025)
Materia
Cryopreservation
Linoleic Acid
In Vitro Experimentation
Oocytes
Bovinae
In Vitro Maturation
Criopreservación
Acido Linoléico
Experimentación in Vitro
Ovocito
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
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spelling Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrificationRios, Glenda LauraSuqueli García, María FlorenciaManrique, Rodrigo J.Buschiazzo, JorgelinaCryopreservationLinoleic AcidIn Vitro ExperimentationOocytesBovinaeIn Vitro MaturationCriopreservaciónAcido LinoléicoExperimentación in VitroOvocitoLong-term conservation of livestock female genetic resources has led to increased interest in oocyte cryopreservation. However, the challenges of preserving large-volume cells and the lack of standardization in vitrification media and cryo-devices for domestic species remain critical areas of ongoing research. This study aimed to assess the impact of in vitro maturation medium composition and vitrification devices on cell death and membrane fusion competence of vitrified bovine oocytes, comparing a chemically undefined versus a synthetic maturation medium supplemented with linoleic acid (LA), and using both tubular (open-pulled straw) and surface vitrification devices (Cryotech®). In vitro maturation of bovine oocytes under serum-free conditions improved post-vitrification survival, particularly by preserving membrane integrity. Additionally, oocytes matured in a synthetic medium containing epidermal growth factor and hyaluronic acid, and vitrified using a surface device, exhibited increased viability and reduced caspase activation. Supplementation of this medium with 43 μM LA did not compromise oocyte viability or apoptotic status. In contrast, higher concentrations (100 μM) induced significant apoptosis and disrupted membrane integrity, impairing tolerance to cryoprotectant agents. We also identified two distinct localization patterns of the transcription factor OCT4 in matured oocytes, with vitrification promoting the diffuse pattern. Supplementation with LA was insufficient to mitigate the effects of vitrification on OCT4 distribution. Unlike 43 μM, maturation with 100 μM LA increased the proportion of oocytes exhibiting the diffuse OCT4 pattern following exposure to cryoprotectant agents under non-vitrified conditions. Zona pellucida-free oocytes co-incubated with sperm demonstrated that 43 μM LA better preserved the original sperm adhesion and fusion pattern post-vitrification, although fertilization rate was not improved. These findings demonstrate that optimized maturation conditions—comprising serum-free media, membrane fluidity-modulating lipids (43 μM LA), and the use of surface cryo-devices—significantly enhance oocyte quality, post-warming survival, and fusion competence, all of which are critical for improving fertilization outcomes following vitrification.EEA BalcarceFil: Ríos, Glenda Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Suqueli García, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Suqueli García, María Florencia. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; ArgentinaFil: Manrique, Rodrigo J. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Buschiazzo, Jorgelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFrontiers Media2025-07-07T10:54:26Z2025-07-07T10:54:26Z2025-07-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/22921https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2025.1628947/full22971769https://doi.org/10.3389/fvets.2025.1628947Frontiers in Veterinary Science 12: 1-14 (03 July 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2023-PD-L01-I112, Biotecnologías reproductivas y plataforma de edición génica para animales de interés zootécnico.info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-04T09:51:10Zoai:localhost:20.500.12123/22921instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:51:10.857INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
title Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
spellingShingle Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
Rios, Glenda Laura
Cryopreservation
Linoleic Acid
In Vitro Experimentation
Oocytes
Bovinae
In Vitro Maturation
Criopreservación
Acido Linoléico
Experimentación in Vitro
Ovocito
title_short Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
title_full Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
title_fullStr Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
title_full_unstemmed Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
title_sort Optimization of bovine oocyte cryopreservation: membrane fusion competence and cell death of linoleic acid-in vitro matured oocytes subjected to vitrification
dc.creator.none.fl_str_mv Rios, Glenda Laura
Suqueli García, María Florencia
Manrique, Rodrigo J.
Buschiazzo, Jorgelina
author Rios, Glenda Laura
author_facet Rios, Glenda Laura
Suqueli García, María Florencia
Manrique, Rodrigo J.
Buschiazzo, Jorgelina
author_role author
author2 Suqueli García, María Florencia
Manrique, Rodrigo J.
Buschiazzo, Jorgelina
author2_role author
author
author
dc.subject.none.fl_str_mv Cryopreservation
Linoleic Acid
In Vitro Experimentation
Oocytes
Bovinae
In Vitro Maturation
Criopreservación
Acido Linoléico
Experimentación in Vitro
Ovocito
topic Cryopreservation
Linoleic Acid
In Vitro Experimentation
Oocytes
Bovinae
In Vitro Maturation
Criopreservación
Acido Linoléico
Experimentación in Vitro
Ovocito
dc.description.none.fl_txt_mv Long-term conservation of livestock female genetic resources has led to increased interest in oocyte cryopreservation. However, the challenges of preserving large-volume cells and the lack of standardization in vitrification media and cryo-devices for domestic species remain critical areas of ongoing research. This study aimed to assess the impact of in vitro maturation medium composition and vitrification devices on cell death and membrane fusion competence of vitrified bovine oocytes, comparing a chemically undefined versus a synthetic maturation medium supplemented with linoleic acid (LA), and using both tubular (open-pulled straw) and surface vitrification devices (Cryotech®). In vitro maturation of bovine oocytes under serum-free conditions improved post-vitrification survival, particularly by preserving membrane integrity. Additionally, oocytes matured in a synthetic medium containing epidermal growth factor and hyaluronic acid, and vitrified using a surface device, exhibited increased viability and reduced caspase activation. Supplementation of this medium with 43 μM LA did not compromise oocyte viability or apoptotic status. In contrast, higher concentrations (100 μM) induced significant apoptosis and disrupted membrane integrity, impairing tolerance to cryoprotectant agents. We also identified two distinct localization patterns of the transcription factor OCT4 in matured oocytes, with vitrification promoting the diffuse pattern. Supplementation with LA was insufficient to mitigate the effects of vitrification on OCT4 distribution. Unlike 43 μM, maturation with 100 μM LA increased the proportion of oocytes exhibiting the diffuse OCT4 pattern following exposure to cryoprotectant agents under non-vitrified conditions. Zona pellucida-free oocytes co-incubated with sperm demonstrated that 43 μM LA better preserved the original sperm adhesion and fusion pattern post-vitrification, although fertilization rate was not improved. These findings demonstrate that optimized maturation conditions—comprising serum-free media, membrane fluidity-modulating lipids (43 μM LA), and the use of surface cryo-devices—significantly enhance oocyte quality, post-warming survival, and fusion competence, all of which are critical for improving fertilization outcomes following vitrification.
EEA Balcarce
Fil: Ríos, Glenda Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Suqueli García, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Suqueli García, María Florencia. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina
Fil: Manrique, Rodrigo J. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Buschiazzo, Jorgelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
description Long-term conservation of livestock female genetic resources has led to increased interest in oocyte cryopreservation. However, the challenges of preserving large-volume cells and the lack of standardization in vitrification media and cryo-devices for domestic species remain critical areas of ongoing research. This study aimed to assess the impact of in vitro maturation medium composition and vitrification devices on cell death and membrane fusion competence of vitrified bovine oocytes, comparing a chemically undefined versus a synthetic maturation medium supplemented with linoleic acid (LA), and using both tubular (open-pulled straw) and surface vitrification devices (Cryotech®). In vitro maturation of bovine oocytes under serum-free conditions improved post-vitrification survival, particularly by preserving membrane integrity. Additionally, oocytes matured in a synthetic medium containing epidermal growth factor and hyaluronic acid, and vitrified using a surface device, exhibited increased viability and reduced caspase activation. Supplementation of this medium with 43 μM LA did not compromise oocyte viability or apoptotic status. In contrast, higher concentrations (100 μM) induced significant apoptosis and disrupted membrane integrity, impairing tolerance to cryoprotectant agents. We also identified two distinct localization patterns of the transcription factor OCT4 in matured oocytes, with vitrification promoting the diffuse pattern. Supplementation with LA was insufficient to mitigate the effects of vitrification on OCT4 distribution. Unlike 43 μM, maturation with 100 μM LA increased the proportion of oocytes exhibiting the diffuse OCT4 pattern following exposure to cryoprotectant agents under non-vitrified conditions. Zona pellucida-free oocytes co-incubated with sperm demonstrated that 43 μM LA better preserved the original sperm adhesion and fusion pattern post-vitrification, although fertilization rate was not improved. These findings demonstrate that optimized maturation conditions—comprising serum-free media, membrane fluidity-modulating lipids (43 μM LA), and the use of surface cryo-devices—significantly enhance oocyte quality, post-warming survival, and fusion competence, all of which are critical for improving fertilization outcomes following vitrification.
publishDate 2025
dc.date.none.fl_str_mv 2025-07-07T10:54:26Z
2025-07-07T10:54:26Z
2025-07-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/22921
https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2025.1628947/full
22971769
https://doi.org/10.3389/fvets.2025.1628947
url http://hdl.handle.net/20.500.12123/22921
https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2025.1628947/full
https://doi.org/10.3389/fvets.2025.1628947
identifier_str_mv 22971769
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2023-PD-L01-I112, Biotecnologías reproductivas y plataforma de edición génica para animales de interés zootécnico.
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv Frontiers in Veterinary Science 12: 1-14 (03 July 2025)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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