Efecto del uso de medio secuencial humano en la producción de blastocistos de hembra ovis canadensis mexicana por clonación manual

Vázquez-Avendaño, José Roberto; Hernández-Martínez, S. E.; Hernández-Pichardo, José Ernesto; Rivera-Rebolledo, Juan Arturo; Ambríz-García, Demetrio Alonso; Navarro-Maldonado, María del Carmen

doi:10.21829/azm.2017.3321068

Cited by 4 publications

(1 citation statement)

References 37 publications

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“…Prior to and after vitrification, following the methodology of Vazquez-Avendaño et al [19] with some modifications, on day 7 of culture, the number of nuclei present in the blastocysts was determined. To accomplish this, embryos of each group were washed for 2 min in 100 µL of DPBS (Dulbeco Phosphate Buffered Saline In Vitro, SA, Mexico City, Mexico) and fixed for 24 h in 400 µL of 4% paraformaldehyde in the refrigerator.…”

Section: Dapi Staining Of Nucleimentioning

confidence: 99%

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

González-Garzón,

Ramón-Ugalde,

Ambríz-García

et al. 2023

Animals

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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 µM, 0.25 µM, 0.5 µM, and 1 µM 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′,7′-dichlorodihydrofluorescein diacetate) to determine the presence of ROS and with Cell Tracker Blue® 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 < 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.

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Section: Dapi Staining Of Nucleimentioning

confidence: 99%

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

González-Garzón,

Ramón-Ugalde,

Ambríz-García

et al. 2023

Animals

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Developmental dynamics of cloned Mexican bighorn sheep embryos using morphological quality standards

Martínez

Pichardo

Avendaño

et al. 2020

Veterinary Medicine & Sci

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The developmental dynamics of cloned Mexican bighorn sheep ( Ovis canadensis mexicana ) embryos were evaluated based on morphological quality standards. Categories determined by standards were correlated with the embryonic development stage, number of nuclei and viability. The results showed no differences in the blastocyst rate between the experimental (cloned Mexican bighorn sheep embryos) and control (parthenogenetic domestic sheep embryos) groups ( p > .05), while type IV fragmentation was higher in clones ( p < .05). The standards allowed for the identification of embryos that divided at least once or fragmented after 24 hr of culture. The highest percentage of morulae appeared at 96 hr, the final stages of development: nonsegmented, blocked, fragmented and blastocysts appeared at 192 hr. Embryonic quality decreased over time, making 96 hr the ideal time point to predict the final morphological quality of embryos. Nuclear staining of the morulae and blastocysts showed that higher embryo quality was associated with a higher percentage of normal and viable blastomeres. The evaluated criteria allowed for descriptions of the dynamics, stage and quality of cloned Mexican bighorn sheep embryos with a high degree of reliability. In addition, developmental anomalies, including fragmentation, multinucleation and blocking, were identified.

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Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

González-Garzón,

Ramón-Ugalde,

Ambríz-García

et al. 2023

Preprint

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The in vitro production and cryopreservation of mammalian embryos generate 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 improving 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 effect of resveratrol in 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 0µM, 0.25µM, 0.5µM and 1µM during oocyte in vitro maturation (IVM). Matured oocytes were fertilized with thawed ram sperm. Embryos were cultured in sequential media until blastocysts, then were vitrified for 24 h and after heating they were stained with DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) to determine the presence of ROS and with Cell Tracker Blue® 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 increases GSH and decreases ROS production (p<0.05). It is concluded that its use in sheep oocytes during IVM has a beneficial effect on embryos produced by IVF subjected to vitrification, protecting them from oxidative damage by promoting the synthesis of antioxidants.

show abstract

Efecto del uso de medio secuencial humano en la producción de blastocistos de hembra ovis canadensis mexicana por clonación manual

Cited by 4 publications

References 37 publications

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

Developmental dynamics of cloned Mexican bighorn sheep embryos using morphological quality standards

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

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