Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro

Ilustraciones, fotografías, tablas

Autores:
Escobar Escobar, Juan Camilo
Tipo de recurso:
Fecha de publicación:
2021
Institución:
Universidad Nacional de Colombia
Repositorio:
Universidad Nacional de Colombia
Idioma:
spa
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oai:repositorio.unal.edu.co:unal/80380
Acceso en línea:
https://repositorio.unal.edu.co/handle/unal/80380
https://repositorio.unal.edu.co/
Palabra clave:
630 - Agricultura y tecnologías relacionadas::636 - Producción animal
Recursos genéticos
Genetic resources
Criopreservación
Cryopreservation
Desarrollo embrionario
Embryonic development
Resveratrol
Biotecnología
Bovinos Criollos
Atemperado
Estrés Oxidativo
Vitrificación
Rights
openAccess
License
Reconocimiento 4.0 Internacional
id UNACIONAL2_1f36fc27971a93122f8649005c1b451a
oai_identifier_str oai:repositorio.unal.edu.co:unal/80380
network_acronym_str UNACIONAL2
network_name_str Universidad Nacional de Colombia
repository_id_str
dc.title.spa.fl_str_mv Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
dc.title.translated.eng.fl_str_mv Evaluation of the antioxidant effect of Resveratrol on the cryotolerance of bovine embryos of the Hartón del Valle breed produced in vitro
title Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
spellingShingle Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
630 - Agricultura y tecnologías relacionadas::636 - Producción animal
Recursos genéticos
Genetic resources
Criopreservación
Cryopreservation
Desarrollo embrionario
Embryonic development
Resveratrol
Biotecnología
Bovinos Criollos
Atemperado
Estrés Oxidativo
Vitrificación
title_short Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
title_full Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
title_fullStr Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
title_full_unstemmed Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
title_sort Evaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitro
dc.creator.fl_str_mv Escobar Escobar, Juan Camilo
dc.contributor.advisor.none.fl_str_mv Torres Osorio, Viviana
Campos Gaona, Rómulo
dc.contributor.author.none.fl_str_mv Escobar Escobar, Juan Camilo
dc.contributor.educationalvalidator.none.fl_str_mv Maturana Mena, Diana Milena
dc.contributor.researchgroup.spa.fl_str_mv Conservación, mejoramiento y utilización del ganado criollo Hartón del Valle y otros recursos genéticos animales en el suroccidente colombiano
dc.subject.ddc.spa.fl_str_mv 630 - Agricultura y tecnologías relacionadas::636 - Producción animal
topic 630 - Agricultura y tecnologías relacionadas::636 - Producción animal
Recursos genéticos
Genetic resources
Criopreservación
Cryopreservation
Desarrollo embrionario
Embryonic development
Resveratrol
Biotecnología
Bovinos Criollos
Atemperado
Estrés Oxidativo
Vitrificación
dc.subject.agrovoc.none.fl_str_mv Recursos genéticos
Genetic resources
Criopreservación
Cryopreservation
Desarrollo embrionario
Embryonic development
Resveratrol
dc.subject.proposal.spa.fl_str_mv Biotecnología
Bovinos Criollos
Atemperado
Estrés Oxidativo
Vitrificación
description Ilustraciones, fotografías, tablas
publishDate 2021
dc.date.accessioned.none.fl_str_mv 2021-10-05T03:48:29Z
dc.date.available.none.fl_str_mv 2021-10-05T03:48:29Z
dc.date.issued.none.fl_str_mv 2021-07-07
dc.type.spa.fl_str_mv Trabajo de grado - Maestría
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.version.spa.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.coarversion.spa.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.content.spa.fl_str_mv Text
dc.type.redcol.spa.fl_str_mv http://purl.org/redcol/resource_type/TM
status_str publishedVersion
dc.identifier.uri.none.fl_str_mv https://repositorio.unal.edu.co/handle/unal/80380
dc.identifier.instname.spa.fl_str_mv Universidad Nacional de Colombia
dc.identifier.reponame.spa.fl_str_mv Repositorio Institucional Universidad Nacional de Colombia
dc.identifier.repourl.spa.fl_str_mv https://repositorio.unal.edu.co/
url https://repositorio.unal.edu.co/handle/unal/80380
https://repositorio.unal.edu.co/
identifier_str_mv Universidad Nacional de Colombia
Repositorio Institucional Universidad Nacional de Colombia
dc.language.iso.spa.fl_str_mv spa
language spa
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spelling Reconocimiento 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Torres Osorio, Viviana8a0c9bca367e01ac8a0ea640c8eb7685Campos Gaona, Rómulof8816487732e56dd010acaeeb7d6bfdaEscobar Escobar, Juan Camilod31390ddca75eb31bc73e6f3ef93e089Maturana Mena, Diana MilenaConservación, mejoramiento y utilización del ganado criollo Hartón del Valle y otros recursos genéticos animales en el suroccidente colombiano2021-10-05T03:48:29Z2021-10-05T03:48:29Z2021-07-07https://repositorio.unal.edu.co/handle/unal/80380Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/Ilustraciones, fotografías, tablasLa raza Hartón del Valle pertenece al conjunto de razas bovinas criollas colombianas adaptadas a las condiciones del trópico, la cual, ha sido sometida a constantes cruzamientos con razas introducidas, reduciendo la población pura y ubicándola en la categoría de “vulnerable”, es decir, que se está enfrentando a un riesgo de extinción alto. En consecuencia, se buscan alternativas para conservar el material genético de la raza. La criopreservación de embriones se ha convertido en un método altamente utilizado en embriones comerciales, debido a la similitud de supervivencia entre los embriones es frescos y los criopreservados (Shaw et al., 2000). La cualidad más relevante de la criopreservación de embriones, es lograr el almacenamiento en condiciones de bajas temperaturas (-196ºC), tratando de mantener la integridad general del embrión (Rodríguez & Jiménez, 2011). Para lograr esto, es necesario eliminar las dos causas principales de muerte celular asociada con la criopreservación, esto es, la formación de cristales de hielo y las concentraciones letales de solutos, mientras se mantiene la integridad de los orgánulos intracelulares (Edgar & Gook, 2012). La vitrificación es un método ideal para criopreservar oocitos y embriones de mamíferos, debido a las altas tasas de enfriamiento y, el corto tiempo de exposición de las células embrionarias a temperaturas críticas y a los crioprotectores, factores que minimizan los efectos tóxicos y el daño a la membrana de las células embrionarias. Lo anterior, perfila la técnica de vitrificación como una alternativa para criopreservar la variabilidad del material genético bovino, sin embargo, se conoce que tiene algunos efectos nocivos sobre la calidad de los embriones. El objetivo de este estudio fue evaluar el efecto de la suplementación del Resveratrol en los medios de cultivo in vitro (CIV) y de atemperado, sobre el desarrollo embrionario, criotolerancia y estado oxidativo de embriones producidos in vitro. Se realizó un primer estudio utilizando oocitos obtenidos de una planta de faenado, se maduraron y fertilizaron in vitro mediante un proceso estandarizado. Los presuntos cigotos se cultivaron en medio SOF suplementado con 0.5 μM de Resveratrol (CR) y sin Resveratrol (C-). El día 7 post fertilización, se evaluaron las tasas de blastocistos y se vitrificaron usando el método de mínimo volumen. Posteriormente, ambos grupos fueron atemperados con Resveratrol 0.5 μM. (C-VR,CRVR) y sin Resveratrol (C-V-: control, CRV-) y fueron cultivados 48 horas para evaluarlas tasas de reexpansión y eclosión. Finalmente, los embriones se sometieron a una doble tinción para medir los niveles de ROS y contenido de GSH intracelular utilizando la sonda 2,7- Diclorodihidrofluoresceina diacetato (H2DCFDA; Invitrogen®) y la sonda 4-clorometil-6,8-difluoro-7-hidroxicumarina (Cell Tracker Blue; CMF2HC; Invitrogen®) respectivamente. El contenido de GSH fue significativamente más alto (p <0.05) en el grupo CRVR en comparación con el grupo control (129.28 ± 8.46% y 100 ± 5.28%, respectivamente). Basados en estos resultados, se realizó un segundo estudio con oocitos obtenidos de aspiración folicular transvaginal guiada por ultrasonografía en hembras Hartón del Valle. Los oocitos se maduraron y fertilizaron in vitro mediante un proceso estandarizado. Los presuntos cigotos se cultivaron en medio SOF suplementado con 0.5 μM de Resveratrol (CR) y sin Resveratrol (C-). El día 7 post fertilización, se evaluaron las tasas de blastocistos y se vitrificaron usando el método de mínimo volumen. Consecutivamente, el grupo cultivado sin Resveratrol se atempero sin Resveratrol (C-V-: control) y el grupo cultivado con Resveratrol se atempero con Resveratrol (CRVR) y fueron cultivados 48 horas para evaluarlas tasas de reexpansión, eclosión, niveles de ROS y contenido de GSH intracelular. Los resultados mostraron un efecto significativo (p <0,05) del Resveratrol sobre las tasas de reexpansión y eclosión embrionaria comparado al grupo control (reexpansión: CRVR= 96.43  3.57% y C-V-= 60.91  4.18%; eclosión CRVR= 37.50  12.84% y C-V-= 17.05  8.51%). Además de una reducción significativa de los niveles de ROS (p <0,05) con respecto al control (CRVR= 73.15 ± 6.01% y C-V-= 100 ± 10.55%). En conclusión, la suplementación simultánea de 0.5 μM Resveratrol en el medio de cultivo in vitro y de atemperado de embriones Hartón del Valle, mejora la supervivencia, criotolerancia y el estado oxidativo de los embriones es producidos in vitro (Texto tomado de la fuente).The Hartón del Valle breed belongs to the set of Colombian Creole bovine breeds adapted to tropical conditions, which has been subjected to constant crossbreeding with introduced breeds, reducing the pure population and placing it in the category of "vulnerable", or facinga high risk of extinction. Consequently, alternatives are being sought to conserve the genetic material of the breed. Embryo cryopreservation has become a highly used method in commercial embryos, due to the survival similarity between fresh and cryopreserved embryos (Shaw et al., 2000). The most relevant attribute on embryo cryopreservation is to achieve storage in low temperature conditions (-196ºC), trying to maintain the general integrity of the embryo (Rodríguez & Jiménez, 2011). To achieve this, it is necessary to eliminate the two main causes of cell death associated with cryopreservation, that is, the formation of ice crystals and lethal concentrations of solutes, while maintaining the integrity of the intracellular organelles (Edgar & Gook, 2012). Vitrification is an ideal method to cryopreserve oocytes and mammalian embryos, due to the high cooling rates and the short time of exposure of embryonic cells to critical temperatures and to cryoprotectants, factors that minimize toxic effects and damage to the embryonic cell membrane. The foregoing outlines the vitrification technique as an alternative to cryopreserve the variability of the bovine genetic material, however, it is known to have some harmful effects on the quality of the embryos. The objective of this study was to evaluate the effect of resveratrol supplementation in in vitro culture (IVC) and tempering media on embryonic development, cryotolerance and oxidative state of in vitro produced embryos. A first study was carried out using oocytes obtained from a slaughter plant, they were matured and fertilized in vitro using a standardized process. The presumed zygotes were cultured in SOF medium supplemented with 0.5 μM of Resveratrol (CR) and without Resveratrol (C-). On day 7 post fertilization, blastocyst rates were evaluated and vitrified using the minimum volume method. Later, both groups were warmed with Resveratrol 0.5 μM. (C-VR, CRVR) and without Resveratrol (C-V-: control, CRV-) and were cultured for 48 hours to evaluate the re-expansion and hatching rates. Finally, the embryos were subjected to double staining to measure ROS levels and intracellular GSH content using the 2,7- Dichlorodihydrofluorescein diacetate probe (H2DCFDA; Invitrogen®) and the 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin probe (Cell Tracker Blue; CMF2HC; Invitrogen®), respectively. The GSH content was significantly higher (p <0.05) in the CRVR group compared to the control group (129.28 ± 8.46% and 100 ± 5.28%, respectively). Based on these results, a second study was carried out with oocytes obtained from transvaginal follicular aspiration guided by ultrasound in Hartón del Valle females. The oocytes were matured and fertilized in vitro using a standardized process. The presumed zygotes were cultured in SOF medium supplemented with 0.5 μM of Resveratrol (CR) and without Resveratrol (C-). On day 7 post fertilization, blastocyst rates were evaluated and vitrified using the minimum volume method. Consecutively, the group cultured without Resveratrol was warmed without Resveratrol (C-V-: control) and the group cultured with Resveratrol was warmed with Resveratrol (CRVR) and were cultured for 48 hours to evaluate the re-expansion rates, hatching rates, ROS levels and GSH content. The results showed a significant effect (p <0.05) of Resveratrol on re-expansion and embryo hatching rates compared to the control group (re-expansion: CRVR= 96.43 ± 3.57% y C-V-= 60.91± 4.18%; hatching = CRVR= 37.50 ± 12.84% y C-V-= 17.05 ± 8.51%). In addition to a significant reduction in ROS levels (p <0.05) with respect to the control (CRVR = 73.15 ±6.01% and C-V- = 100 ± 10.55%). In conclusion, the simultaneous supplementation of 0.5 μM Resveratrol in the in vitro culture medium and the warming medium of Hartón del Valle embryos improves survival, cryotolerance and the oxidative state of embryos produced in vitro.MaestríaMagister en Ciencias AgrariasSe realizó un primer estudio utilizando oocitos obtenidos de una planta de faenado, se maduraron y fertilizaron in vitro mediante un proceso estandarizado. Los presuntos cigotos se cultivaron en medio SOF suplementado con 0.5 μM de Resveratrol (CR) y sin Resveratrol (C-). El día 7 post fertilización, se evaluaron las tasas de blastocistos y se vitrificaron usando el método de mínimo volumen. Posteriormente, ambos grupos fueron atemperados con Resveratrol 0.5 μM. (C-VR, CRVR) y sin Resveratrol (C-V-: control, CRV-) y fueron cultivados 48 horas para evaluarlas tasas de reexpansión y eclosión. Finalmente, los embriones se sometieron a una doble tinción para medir los niveles de ROS y contenido de GSH intracelular utilizando la sonda 2,7- Diclorodihidrofluoresceina diacetato (H2DCFDA; Invitrogen®) y la sonda 4-clorometil6,8-difluoro-7-hidroxicumarina (Cell Tracker Blue; CMF2HC; Invitrogen®) respectivamente.Producción animal tropicalxiv, 141 páginas + anexosapplication/pdfspaUniversidad Nacional de ColombiaPalmira - Ciencias Agropecuarias - Maestría en Ciencias AgrariasFacultad de Ciencias AgropecuariasUniversidad Nacional de Colombia - Sede Palmira630 - Agricultura y tecnologías relacionadas::636 - Producción animalRecursos genéticosGenetic resourcesCriopreservaciónCryopreservationDesarrollo embrionarioEmbryonic developmentResveratrolBiotecnologíaBovinos CriollosAtemperadoEstrés OxidativoVitrificaciónEvaluación del efecto antioxidante del Resveratrol sobre la criotolerancia de embriones bovinos de la raza Hartón del Valle producidos in vitroEvaluation of the antioxidant effect of Resveratrol on the cryotolerance of bovine embryos of the Hartón del Valle breed produced in vitroTrabajo de grado - Maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85Texthttp://purl.org/redcol/resource_type/TMValle del Cauca , ColombiaAbdelatty, A.M., Iwaniuk, M.E., Potts, S.B., Gad, A., 2018. 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Theriogenology, New York 85, 688–697.InvestigadoresLICENSElicense.txtlicense.txttext/plain; charset=utf-83964https://repositorio.unal.edu.co/bitstream/unal/80380/1/license.txtcccfe52f796b7c63423298c2d3365fc6MD51ORIGINAL1114823584.2021.pdf1114823584.2021.pdfMaestría en Ciencias Agrariasapplication/pdf3690727https://repositorio.unal.edu.co/bitstream/unal/80380/3/1114823584.2021.pdfb03ce24efc170c9d28df7ca88d2220f9MD53THUMBNAIL1114823584.2021.pdf.jpg1114823584.2021.pdf.jpgGenerated Thumbnailimage/jpeg5719https://repositorio.unal.edu.co/bitstream/unal/80380/4/1114823584.2021.pdf.jpg8f3376e79427efa9d73f84f9c8c9d87aMD54unal/80380oai:repositorio.unal.edu.co:unal/803802023-07-23 23:04:17.864Repositorio Institucional Universidad Nacional de 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GVyZWNob3MgZGUgYXV0b3IgcXVlIGNvbmxsZXZlIGxhIGRpc3RyaWJ1Y2nDs24gZGUgZXN0b3MgYXJjaGl2b3MgeSBtZXRhZGF0b3MuCkFsIGhhY2VyIGNsaWMgZW4gZWwgc2lndWllbnRlIGJvdMOzbiwgdXN0ZWQgaW5kaWNhIHF1ZSBlc3TDoSBkZSBhY3VlcmRvIGNvbiBlc3RvcyB0w6lybWlub3MuCgpVTklWRVJTSURBRCBOQUNJT05BTCBERSBDT0xPTUJJQSAtIMOabHRpbWEgbW9kaWZpY2FjacOzbiAyNy8yMC8yMDIwCg==

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