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dc.contributor.author
Pereira, Blasa C.
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dc.contributor.author
Ortiz, Isabel
dc.contributor.author
Dorado, Jesús
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dc.contributor.author
Diaz Jimenez, Maria
dc.contributor.author
Consuegra, Cesar
dc.contributor.author
Demyda Peyrás, Sebastián
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dc.contributor.author
Hidalgo, Manuel
dc.date.available
2021-08-20T01:33:18Z
dc.date.issued
2020-07
dc.identifier.citation
Pereira, Blasa C.; Ortiz, Isabel; Dorado, Jesús; Diaz Jimenez, Maria; Consuegra, Cesar; et al.; The Effect of different vitrification and staining protocols on the visibility of the nuclear maturation stage of equine oocytes; Elsevier Science Inc.; Journal of Equine Veterinary Science; 90; 103021; 7-2020; 1-5
dc.identifier.issn
0737-0806
dc.identifier.uri
http://hdl.handle.net/11336/138561
dc.description.abstract
In this study, we compared two staining protocols assessing the nuclear chromatin stage of equine oocytes after vitrification using permeable and nonpermeable cryoprotectants. Slaughterhouse-derived oocytes (n = 155) were obtained from a total of 32 mares and in vitro matured in M199 medium for 42 hours at 38.5°C in 5% CO2. In the first experiment, two concentrations of Hoechst 33342 (HO) were tested (10 μg/mL; P1 and 2.5 μg/mL; P2) combined with 50 μg/mL of propidium iodide as staining protocols to evaluate the visibility of matured oocytes (n = 44). In the second experiment, 111 oocytes were evaluated using the staining protocol P2, before (C, control) and after vitrification following a two-step conventional protocol with (15% dimethyl sulfoxide, 15% ethylene glycol, and 0.5 M sucrose; V1) or without (1 M sucrose; V2) using permeable cryoprotectants. Our results showed that P2 provided a higher percentage of oocytes with outstanding visibility of the nuclear chromatin stage (52.17%; P < .05) in comparison with P1 (19.04%). In the second experiment, no cryoprotectant-free vitrified oocytes reached the metaphase II maturation stage. This result was significantly lower (P < .05) than conventional vitrification (15.38%) and both lower in comparison with the nonvitrified control group (42.11%). In conclusion, permeable cryoprotectant-free vitrification of equine oocytes obtained poor results and therefore cannot be considered an alternative to vitrification using permeable cryoprotectants. In addition, a staining protocol with a low concentration of HO is recommended to evaluate the nuclear chromatin stage of equine oocytes after in vitro maturation.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier Science Inc.
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dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
CRYOPRESERVATION
dc.subject
CRYOPROTECTANT
dc.subject
HOECHST 33342
dc.subject
MARE
dc.subject
MEIOTIC COMPETENCE
dc.subject.classification
Tecnología GM, clonación de ganado, selección asistida, diagnósticos, tecnología de producción de biomasa, etc.
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dc.subject.classification
Biotecnología Agropecuaria
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dc.subject.classification
CIENCIAS AGRÍCOLAS
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dc.title
The Effect of different vitrification and staining protocols on the visibility of the nuclear maturation stage of equine oocytes
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-07-30T18:26:37Z
dc.journal.volume
90
dc.journal.number
103021
dc.journal.pagination
1-5
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Pereira, Blasa C.. Universidad de Córdoba; España
dc.description.fil
Fil: Ortiz, Isabel. Universidad de Córdoba; España
dc.description.fil
Fil: Dorado, Jesús. Universidad de Córdoba; España
dc.description.fil
Fil: Diaz Jimenez, Maria. Universidad de Córdoba; España
dc.description.fil
Fil: Consuegra, Cesar. Universidad de Córdoba; España
dc.description.fil
Fil: Demyda Peyrás, Sebastián. Universidad Nacional de La Plata; Argentina. Universidad de Córdoba; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina
dc.description.fil
Fil: Hidalgo, Manuel. Universidad de Córdoba; España
dc.journal.title
Journal of Equine Veterinary Science
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S073708062030112X?via%3Dihub
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.jevs.2020.103021
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