Effect of vitrification and beta-mercaptoethanol on reactive oxygen species activity and in vitro development of oocytes vitrified before or after in vitro fertilization

Gupta, Mukesh Kumar; Uhm, Sang Jun; Lee, Hoon Taek

doi:10.1016/j.fertnstert.2010.01.043

Cited by 159 publications

(91 citation statements)

References 44 publications

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“…High O2 tension (20%) induced ROS generation in porcine embryos, but the detrimental effects of ROS were overcome by the combination of GSH or β-ME with cysteine as in previous studies [7,43,44].…”

Section: Additional Effects Of Glutathione and β-Mercaptoethanol Treamentioning

confidence: 73%

Synergistic Effects of Glutathione and .BETA.-Mercaptoethanol Treatment During In Vitro Maturation of Porcine Oocytes on Early Embryonic Development in a Culture System Supplemented with L-cysteine

et al. 2010

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Abstract. Various methods have been used to remove reactive oxygen species (ROS) generated from in vitro culture (IVC) conditions that can cause cell injury or death, including the application of low oxygen (O2) tension and the addition of antioxidants. The beneficial effects of antioxidants and O2 tension on IVC of porcine embryos, however, are controversial among researchers. In this study, we sought to determine the effects and optimal concentrations of antioxidants for the development of porcine embryos in an IVC system. Specifically, we examined the synergistic effects of antioxidants on development to the blastocyst stage in a culture system supplemented with L-cysteine during IVM. Of the antioxidants tested (melatonin, glutathione (GSH), β-mercaptoethanol (β-ME), N-acetylcysteine (NAC) and dithiothreitol (DTT)), addition of GSH (1 mM) or β-ME (25 μM) significantly increased development to the blastocyst stage compared with the controls without antioxidant treatment (22.2 ± 4.2% for 1 mM GSH, 25.9 ± 2.2% for 25 μM β-ME and 12-13% for the control, P<0.05). In addition, the mean cell number per blastocyst was increased by approximately 1.7-fold in the presence of GSH or β -ME. These GSH-and β-ME-induced increases in development to the blastocyst stage and total cell number, however, were not mimicked by melatonin, NAC or DTT, all of which are ROS scavengers. The combination of GSH or β-ME with L-cysteine significantly reduced high O2 tension-induced ROS production (P<0.05). These results suggest that a combination of 1 mM GSH or 25 μM β-ME with 1 mM L-cysteine could be used for production of high quality porcine blastocysts in IVC systems. Key words: Antioxidant system, Embryo, Oxidative stress, Pig (J. Reprod. Dev. 56: [575][576][577][578][579][580][581][582] 2010) he academic and industrial value of porcine embryonic technology is very high, as this technology allows for production of transgenic porcine embryos with disease-resistance genes, for xenogenic organ transplantation and for preservation of superior genotypes that are at risk of extinction. Obtaining many high-quality embryos, however, is a prerequisite for improvement of embryo technology. A large number of immature oocytes can be collected from the ovaries of slaughtered pigs, and these oocytes can then be matured, fertilized and cultured in vitro. At present, the quality of in vitro produced (IVP) porcine embryos is considerably lower than that of in vivo-derived embryos, although many investigators have attempted to improve in vitro culture systems to help produce higher quality porcine embryos.Earlier studies have shown that in vitro cultured (IVC) embryos cannot develop normally, as the oxygen (O2) tension in vitro is higher than that in the oviduct. This high O2 tension induces excessive production of reactive oxygen species (ROS), which leads to oxidative stress and impedes oocyte maturation and embryonic development [1,2]. To remove the ROS generated from IVC conditions, various methods have been used such as the application of low O2...

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Section: Additional Effects Of Glutathione and β-Mercaptoethanol Treamentioning

confidence: 73%

Synergistic Effects of Glutathione and .BETA.-Mercaptoethanol Treatment During In Vitro Maturation of Porcine Oocytes on Early Embryonic Development in a Culture System Supplemented with L-cysteine

et al. 2010

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“…Similarly, in our laboratory, we have succeeded in the generation of blastocysts from vitrified M-II stage oocytes by parthenogenetic activation [25], whereas IVF of the M-II stage oocytes vitrified by the same method did not result in blastocyst development [27]. The primary problem preventing embryo development from vitrified porcine M-II stage oocytes is the reduction of their ability to be fertilized by spermatozoa in vitro which has been experienced in several studies [12, 24, 25]. Hardening of the zona pellucida caused by the cryopreservation process has been observed in mammalian oocytes [44,45] which may provide a reasonable explanation for the reduced penetration rates.…”

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confidence: 99%

Factors Affecting Cryopreservation of Porcine Oocytes

2012

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“…Com relação à criotolerância, aparentemente, o BM é mais benéfico se adicionado no cultivo, logo após o processo de criopreservação. Nedambale et al (34) demonstraram aumento na sobrevivência, eclosão e no número de células de blastocistos bovinos, com a adição de BM no meio de cultivo após a vitrificação, o que também foi observado por GUPTA et al (35) . É possível que o estresse produzido pelo processo de vitrificação, sobre os blastocistos, seja prevenido pela subsequente adição de BM.…”

Section: Resultsunclassified

CRIOTOLERÂNCIA DE OÓCITOS E EMBRIÕES BOVINOS MATURADOS COM LÍQUIDO FOLICULAR E/Ou Β-Mercaptoetanol

et al. 2015

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ResumoFoi avaliada a criotolerância de oócitos e embriões bovinos maturados com adição de líquido folicular (LF) e/ou β-mercaptoetanol (BM). Após vitrificação, os oócitos foram maturados em: TCM-199 (controle); BM (24h TCM-199+100µM BM); LF (6h em LF+18h TCM-199) e LF+BM (6h LF+18h TCM-199+100µM BM). Não houve diferença (p>0,05) nas taxas de blastocistos dos tratamentos TCM (6,4%), BM (4,0%) e LF (3,4%). A eclosão e densidade celular dos embriões eclodidos não diferiram (p>0,05) nos tratamentos. No Experimento 2 blastocistos expandidos (Bx) obtidos em D7 ou D8 foram vitrificados, avaliando-se sua reexpansão e eclosão. A reexpansão foi semelhante (p>0,05), sendo observado comportamento distinto na eclosão entre Bx D7 e D8. Nos Bx D7 houve maior eclosão no controle (TCM-54,2%) em relação ao BM (40,32%) e LF+BM (33,89%). Os Bx D8 apresentaram menor eclosão no controle (TCM) em relação aos Bx D7. Nos tratamentos BM, LF e LF+BM a eclosão foi semelhante para Bx D7 ou D8. A maturação com adição de LF e/ou BM não melhora a criotolerância de oócitos imaturos e embriões PIV. Blastocistos expandidos precoces (D7) são mais criotolerantes e apresentam um comportamento distinto à adição de LF e BM, em relação aos tardios (D8). Palavras-chave: aditivos; criotolerância; embrião PIV; maturação; vitrificação. AbstractCryotolerance of bovine oocytes and embryos maturated with addition of follicular fluid (LF) and β-

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Effect of vitrification and beta-mercaptoethanol on reactive oxygen species activity and in vitro development of oocytes vitrified before or after in vitro fertilization

Cited by 159 publications

References 44 publications

Synergistic Effects of Glutathione and .BETA.-Mercaptoethanol Treatment During In Vitro Maturation of Porcine Oocytes on Early Embryonic Development in a Culture System Supplemented with L-cysteine

Synergistic Effects of Glutathione and .BETA.-Mercaptoethanol Treatment During In Vitro Maturation of Porcine Oocytes on Early Embryonic Development in a Culture System Supplemented with L-cysteine

Factors Affecting Cryopreservation of Porcine Oocytes

CRIOTOLERÂNCIA DE OÓCITOS E EMBRIÕES BOVINOS MATURADOS COM LÍQUIDO FOLICULAR E/Ou Β-Mercaptoetanol

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