{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:31:18Z","timestamp":1787027478866,"version":"build-2736575974"},"reference-count":51,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2023,11,21]],"date-time":"2023-11-21T00:00:00Z","timestamp":1700524800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The in vitro production and cryopreservation of mammalian embryos generates reactive oxygen species (ROS) due to conditions of the system that can overcome their antioxidant protection. Resveratrol is an antioxidant used in in vitro systems to improve blastocyst rates, but its effect on antioxidant enzymes such as glutathione (GSH) in embryos produced by in vitro fertilization (IVF) after vitrification has not been reported. The objective of this study was to evaluate the effects of resveratrol on the in vitro maturation medium (IVM) of sheep oocytes (Ovis aries) on the levels of ROS and GSH in embryos produced by IVF subjected to vitrification. Resveratrol was added at 0 \u00b5M, 0.25 \u00b5M, 0.5 \u00b5M, and 1 \u00b5M during oocyte in vitro maturation (IVM). Matured oocytes were fertilized with thawed ram sperm. Embryos were cultured in sequential media until blastocysts, were then vitrified for 24 h, and, after heating, they were stained with DCFH-DA (2\u2032,7\u2032-dichlorodihydrofluorescein diacetate) to determine the presence of ROS and with Cell Tracker Blue\u00ae for the presence of GSH. The quantitative values of ROS and GSH were obtained through the Image J image processor. The results showed that resveratrol increased GSH and decreased ROS production (p &lt; 0.05) in a dose-dependent manner. It is concluded that its use in sheep oocytes during IVM has a beneficial effect on embryos produced by IVF subjected to vitrification by decreasing reactive oxygen species levels and facilitating the generation of embryo antioxidant enzymes like glutathione.<\/jats:p>","DOI":"10.3390\/ani13233602","type":"journal-article","created":{"date-parts":[[2023,11,21]],"date-time":"2023-11-21T12:12:13Z","timestamp":1700568733000},"page":"3602","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos"],"prefix":"10.3390","volume":"13","author":[{"given":"Andrea Carolina","family":"Gonz\u00e1lez-Garz\u00f3n","sequence":"first","affiliation":[{"name":"Division of Biological and Health Sciences, Universidad Aut\u00f3noma Metropolitana Unidad Iztapalapa, Mexico City 09310, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio Porfirio","family":"Ram\u00f3n-Ugalde","sequence":"additional","affiliation":[{"name":"Division of Posgraduate Studies and Research, Tecnol\u00f3gico Nacional de M\u00e9xico Campus Conkal, Yucat\u00e1n 97345, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Demetrio Alonso","family":"Ambr\u00edz-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Biology of Reproduction, Universidad Aut\u00f3noma Metropolitana Unidad Iztapalapa, M\u00e9xico City 09310, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Roberto","family":"Vazquez-Avenda\u00f1o","sequence":"additional","affiliation":[{"name":"Department of Biology of Reproduction, Universidad Aut\u00f3noma Metropolitana Unidad Iztapalapa, M\u00e9xico City 09310, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Ernesto","family":"Hern\u00e1ndez-Pichardo","sequence":"additional","affiliation":[{"name":"Department of Agriculture and Animal Production, Universidad Aut\u00f3noma Metropolitana Unidad Xochimilco, M\u00e9xico City 04960, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Luis","family":"Rodr\u00edguez-Suastegui","sequence":"additional","affiliation":[{"name":"Department of Agriculture and Animal Production, Universidad Aut\u00f3noma Metropolitana Unidad Xochimilco, M\u00e9xico City 04960, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7213-9034","authenticated-orcid":false,"given":"C\u00e9sar","family":"Cortez-Romero","sequence":"additional","affiliation":[{"name":"Campus San Luis Potos\u00ed, Colegio de Postgraduados, Salinas de Hidalgo 78622, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7858-7038","authenticated-orcid":false,"given":"Mar\u00eda del Carmen","family":"Navarro-Maldonado","sequence":"additional","affiliation":[{"name":"Department of Biology of Reproduction, Universidad Aut\u00f3noma Metropolitana Unidad Iztapalapa, M\u00e9xico City 09310, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,21]]},"reference":[{"key":"ref_1","first-page":"79","article-title":"Situaci\u00f3n actual y perspectivas de la ganader\u00eda de bovinos criollos en Am\u00e9rica Latina","volume":"29","year":"2021","journal-title":"Arch. Latinoam. Prod. Anim."},{"key":"ref_2","first-page":"9","article-title":"Desarrollo de m\u00f3rulas de ovino en medio simple o secuencial: Relaci\u00f3n entre evaluaci\u00f3n morfol\u00f3gica y viabilidad embrionaria","volume":"39","year":"2017","journal-title":"Rev. Salud Anim."},{"key":"ref_3","first-page":"1","article-title":"Effect of resveratrol on the in vitro maturation of ovine (Ovis aries) oocytes and the subsequent development of handmade cloned embryos","volume":"5","year":"2019","journal-title":"Vet. M\u00e9x."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/s10815-013-0116-9","article-title":"Resveratrol treatment during goat oocytes maturation enhances developmental competence of parthenogenetic and hand-made cloned blastocysts by modulating intracellular glutathione level and embryonic gene expression","volume":"31","author":"Mukherjee","year":"2014","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"104378","DOI":"10.1016\/j.livsci.2020.104378","article-title":"In vitro maturation medium supplementation with resveratrol improves cumulus cell expansion and developmental competence of Sanjabi sheep oocytes","volume":"243","author":"Zabihi","year":"2021","journal-title":"Livest. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.cryobiol.2019.05.008","article-title":"Effect of resveratrol on vitrified in vitro produced bovine embryos: Recovering the initial quality","volume":"89","author":"Gaviria","year":"2019","journal-title":"Cryobiology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.theriogenology.2012.01.024","article-title":"The effects of resveratrol on porcine oocyte in vitro maturation and subsequent embryonic development after parthenogenetic activation and in vitro fertilization","volume":"78","author":"Kwak","year":"2012","journal-title":"Theriogenology"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Abdul Rahman, N.-S., Mohamed Noor Khan, N.-A., Eshak, Z., Sarbandi, M.-S., Mohammad Kamal, A.-A., Abd Malek, M., Abdullah, F., Abdullah, M.A., and Othman, F. (2022). Exogenous L-Glutathione Improves Vitrification Outcomes in Murine Preimplantation Embryos. Antioxidants, 11.","DOI":"10.3390\/antiox11112100"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Mart\u00ednez, T., Vendrell-Flotats, M., Mart\u00ednez-Rodero, I., Ord\u00f3\u00f1ez-Le\u00f3n, E.A., \u00c1lvarez-Rodr\u00edguez, M., L\u00f3pez-B\u00e9jar, M., Yeste, M., and Mogas, T. (2020). Glutathione Ethyl Ester Protects In Vitro-Maturing Bovine Oocytes against Oxidative Stress Induced by Subsequent Vitrification\/Warming. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21207547"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"871289","DOI":"10.3389\/fvets.2022.871289","article-title":"Astaxanthin supplementation improves the subsequent developmental competence of vitrified porcine zygotes","volume":"9","author":"Xiang","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"ref_11","unstructured":"Giraldo Giraldo, J.J. (2011). Efecto del Crioprotector Dimetilformamida Sobre la Viabilidad de Embriones Bovinos Producidos In Vitro. [Master\u2019s Thesis, Universidad Nacional de Colombia, Facultad de Ciencias, Sede Medell\u00edn]."},{"key":"ref_12","unstructured":"Asociaci\u00f3n para el Estudio de la Biolog\u00eda de la Reproducci\u00f3n (ASEBIR) (2015). Criterios ASEBIR de Valoraci\u00f3n Morfol\u00f3gica de Oocitos, Embriones Tempranos y Blastocistos Humanos, Cuadernos de Embriolog\u00eda Cl\u00ednica. [3rd ed.]."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.4067\/S0717-95022017000401337","article-title":"Comportamiento del Semen Bovino Sexado Congelado-Descongelado en Fertilizaci\u00f3n in vitro (FIV) Capacitado Mediante BO en dos Concentraciones versus Percoll","volume":"35","author":"Filipiak","year":"2017","journal-title":"Int. J. Morphol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e17138","DOI":"10.15381\/rivep.v31i3.17138","article-title":"Aspectos esenciales sobre las t\u00e9cnicas de fertilizaci\u00f3n in vitro en bovinos","volume":"31","author":"Vasquez","year":"2020","journal-title":"Rev. Investig. Vet. Per\u00fa"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1071\/RD14024","article-title":"Open pulled straw vitrification and slow freezing of sheep IVF embryos using different cryoprotectants","volume":"27","author":"Bhat","year":"2015","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/S0378-4320(00)00097-X","article-title":"Vitrification of the oocytes and embryos of domestic animals","volume":"60\u201361","author":"Vajta","year":"2000","journal-title":"Anim. Reprod. Sci."},{"key":"ref_17","unstructured":"Sosa, F., and Hansen, P.J. (2019). Procedures for In Vitro Production of Bovine Embryos, University of Florida."},{"key":"ref_18","unstructured":"Gonz\u00e1lez Mendoza, D.F. (2020). Efecto del Protocolo de Vitrificaci\u00f3n y Sistemas de Empaque Sobre la Tasa de Supervivencia de Embriones Ovinos Obtenidos In Vivo. [Master\u2019s Thesis, Universidad Nacional de Colombia, Facultad de Medicina Veterinaria y de Zootecnia, Programas de Posgrados]."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"328","DOI":"10.21829\/azm.2017.3321068","article-title":"Efecto del uso de medio secuencial humano en la producci\u00f3n de blastocistos de hembra ovis canadensis mexicana por clonaci\u00f3n manual","volume":"33","year":"2017","journal-title":"Acta Zool. Mex."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1093\/humupd\/7.2.175","article-title":"Oxidative stress y protecci\u00f3n contra especies reactivas de ox\u00edgeno en el embri\u00f3n preimplantacional y su entorno","volume":"7","author":"Guerin","year":"2001","journal-title":"Hum. Reprod. Update"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e17557","DOI":"10.15381\/rivep.v31i1.17557","article-title":"Efecto de la adici\u00f3n de melatonina en el medio de maduraci\u00f3n y\/o vitrificaci\u00f3n de ovocitos sobre la producci\u00f3n in vitro de embriones bovinos","volume":"31","author":"Argudo","year":"2020","journal-title":"Rev. Investig. Vet. Per\u00fa"},{"key":"ref_22","unstructured":"Soberano, M.A. (2011). Participaci\u00f3n de Compuestos Tiol en el Medio de Maduraci\u00f3n de Ovocitos de Cabra y su Efecto Sobre la Producci\u00f3n de Embriones In Vitro. [Ph.D. Thesis, Universidad Michoacana de San Nicol\u00e1s de Hidalgo, Doctorado en Biotecnolog\u00eda Molecular Agropecuaria]."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1590\/1678-4162-10323","article-title":"La actividad antioxidante del extracto oleoso obtenido de Lippia origanoides mejora la calidad de los embriones bovinos producidos in vitro","volume":"71","author":"Sollecito","year":"2019","journal-title":"Arq. Bras. Med. Vet. Zootec."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1038\/nsmb.1821","article-title":"Repressive and active histone methylation mark distinct promoters in human and mouse spermatozoa","volume":"17","author":"Brykczynska","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_25","first-page":"8213071","article-title":"Sperm oxidative stress is detrimental to embryo development: A dose-dependent study model and a new and more sensitive oxidative status evaluation","volume":"2016","author":"Siqueira","year":"2015","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Selivanov, V.A., Votyakova, T.V., Pivtoraiko, V.N., Zeak, J., Sukhomlin, T., Trucco, M., Roca, J., and Cascante, M. (2011). Reactive oxygen species production by forward and reverse electron fluxes in the mitochondrial respiratory chain. PLoS Comput. Biol., 7.","DOI":"10.1371\/journal.pcbi.1001115"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1073\/pnas.94.2.514","article-title":"Mitochondrial DNA damage is more extensive and persists longer than nuclear DNA damage in human cells following oxidative stress","volume":"94","author":"Yakes","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/35041687","article-title":"Oxidants, oxidative stress and the biology of ageing","volume":"408","author":"Finkel","year":"2000","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1093\/ije\/dyr232","article-title":"Epigenetics, epidemiology and mito-chondrial DNA diseases","volume":"41","author":"Chinnery","year":"2012","journal-title":"Int. J. Epidemiol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Han, Y., and Chen, J.Z. (2013). Oxidative stress induces mitochondrial dna damage and cytotoxicity through independent mechanisms in human cancer cells. BioMed Res. Int., 2013.","DOI":"10.1155\/2013\/825065"},{"key":"ref_31","first-page":"2003","article-title":"Oxidative stress, mitochondrial damage and neurodegenerative diseases","volume":"8","author":"Guo","year":"2013","journal-title":"Neural Regen. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1006\/scdb.2001.0282","article-title":"Reactive oxygen species and mitochondrial diseases","volume":"12","author":"Kirkinezos","year":"2001","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"864","DOI":"10.1016\/S1472-6483(10)60038-7","article-title":"Podr\u00eda el estr\u00e9s oxidativo influir en la maduraci\u00f3n in vitro de los ovocitos?","volume":"18","author":"Combelles","year":"2009","journal-title":"Reprod. Biomed. Online"},{"key":"ref_34","first-page":"64","article-title":"Maduraci\u00f3n in vitro de ovocitos bovinos: Efectos beneficiosos de la cisteamina","volume":"7","author":"Asiye","year":"2018","journal-title":"J. Dairy Vet. Anim. Res."},{"key":"ref_35","first-page":"310","article-title":"Alpha-tocopherol and Ascorbic Acid Combinations Influence the Maturation of Sheep Oocytes","volume":"45","author":"Miclea","year":"2012","journal-title":"Sci. Pap. Anim. Sci. Biotechnol."},{"key":"ref_36","first-page":"1","article-title":"Assisted Reproductive Technology Surveillance\u2014United States","volume":"67","author":"Sunderam","year":"2015","journal-title":"Surveill. Summ."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2874283","DOI":"10.1155\/2017\/2874283","article-title":"Pluripotent stem cell metabolism and mitochondria: Beyond ATP","volume":"2017","author":"Lees","year":"2017","journal-title":"Stem Cells Int."},{"key":"ref_38","unstructured":"Vargas Reyes, J.N. (2013). Efecto de las T\u00e9cnicas de Congelaci\u00f3n Lenta y Vitrificaci\u00f3n con Etilenglicol Sobre la Calidad Poscriopreservaci\u00f3n de Embriones Bovinos Producidos In Vitro. [Master\u2019s Thesis, Maestr\u00eda en Ciencias Veterinarias Universidad de La Salle, Facultad de Ciencias Agropecuarias]. Available online: https:\/\/ciencia.lasalle.edu.co\/maest_ciencias_veterinarias\/10."},{"key":"ref_39","first-page":"e2764","article-title":"Cryopreservation of Preimplantation Embryos of Cattle, Sheep, and Goats","volume":"54","author":"Youngs","year":"2011","journal-title":"J. Vis. Exp."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"920257","DOI":"10.1155\/2013\/920257","article-title":"Effects of resveratrol on vitrified porcine oocytes","volume":"2013","author":"Giaretta","year":"2013","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.anireprosci.2014.09.018","article-title":"Effect of resveratrol supplementation during culture on the quality and cryotolerance of bovine in vitro produced embryos","volume":"151","author":"Salzano","year":"2014","journal-title":"Anim. Reprod. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s12013-009-9054-7","article-title":"Antioxidant activity of sulfur and selenium: A review of reactive oxygen species scavenging, glutathione peroxidase, and metal-binding antioxidant mechanisms","volume":"55","author":"Battin","year":"2009","journal-title":"Cell Biochem. Biophys."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.1016\/S0093-691X(02)00643-X","article-title":"Effect of glutathione synthesis stimulation during in vitro maturation of ovine oocytes on embryo development and intracellular peroxide content","volume":"57","author":"Gasparrini","year":"2002","journal-title":"Theriogenology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1002\/mrd.10214","article-title":"Glutathione (GSH) concentrations vary with the cell cycle in maturing hamster oocytes, zygotes, and pre-implantation stage embryos","volume":"64","author":"Zuelke","year":"2003","journal-title":"Mol. Reprod. Dev."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1002\/(SICI)1098-2795(199612)45:4<451::AID-MRD7>3.0.CO;2-Q","article-title":"Stimulation of glutathione synthesis of in vitro matured bovine oocytes and its effect on embryo development and freezability","volume":"45","author":"Furnus","year":"1996","journal-title":"Mol. Reprod. Dev."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1095\/biolreprod57.6.1420","article-title":"Glutathione Synthesis during in vitro maturation of bovine oocytes: Role of cumulus cells","volume":"57","author":"Furnus","year":"1997","journal-title":"Biol. Reprod."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1002\/vms3.242","article-title":"Developmental dynamics of cloned Mexican bighorn sheep embryos using morphological quality standards","volume":"6","year":"2020","journal-title":"Vet. Med. Sci."},{"key":"ref_48","first-page":"e87","article-title":"In vitro embryo production from ewes at different physiological stages","volume":"23","year":"2022","journal-title":"J. Vet. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"22629","DOI":"10.1038\/s41598-021-02000-z","article-title":"Controlled spermatozoa\u2013oocyte interaction improves embryo quality in sheep","volume":"11","author":"Anzalone","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Mastrorocco, A., Cacopardo, L., Lamanna, D., Temerario, L., Brunetti, G., Carluccio, A., Robbe, D., and Dell\u2019Aquila, M.E. (2021). Bioengineering approaches to improve in vitro performance of prepubertal lamb oocytes. Cells, 10.","DOI":"10.3390\/cells10061458"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1111\/rda.14088","article-title":"Supplementation of freezing medium with encapsulated or free glutathione during cryopreservation of bull sperm","volume":"57","author":"Nadri","year":"2022","journal-title":"Reprod. Domest. Anim. Zuchthyg."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/23\/3602\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:26:32Z","timestamp":1760131592000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/23\/3602"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,21]]},"references-count":51,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["ani13233602"],"URL":"https:\/\/doi.org\/10.3390\/ani13233602","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,21]]}}}