Genotoxicity assessment of mouse oocytes by comet assay before vitrification and after warming with three vitrification protocols

Berthelot-Ricou, A.; Perrin, Jeanne; Giorgio, Carole Di; Méo, Michel De; Botta, A.; Courbière, Blandine

doi:10.1016/j.fertnstert.2013.05.025

Cited by 21 publications

(11 citation statements)

References 34 publications

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“…Though, impact of vitrification on the DNA of GV oocytes is not elucidated, a significant rate of DNA fragmentation was found in bovine vitrified mature oocytes regardless of the cryopreservation method [Stachowiak et al 2009]. Similarly, vitrification protocols induced significant DNA damage when applied to mouse mature oocytes, both before cooling and after warming [Berthelot-Ricou et al 2013]. In addition, an earlier study has demonstrated that the process of vitrification results in significant degradation of a large part of intact RNA which was not observed by conventional slow freezing [Isachenko et al 2007].…”

Section: Discussionmentioning

confidence: 99%

Ovarian tissue vitrification is more efficient than slow freezing in protecting oocyte and granulosa cell DNA integrity

Mathias

D’Souza

Uppangala

et al. 2014

Systems Biology in Reproductive Medicine

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Ovarian tissue cryopreservation is the primary treatment modality currently available to women at risk of losing their ovarian function due to cytotoxic therapy. However, the impact of these techniques on the oocyte DNA integrity is not elucidated. Here we have investigated the effect of vitrification and conventional slow freezing of eight week old Swiss albino mouse ovarian tissues on the oocyte and granulosa cell DNA integrity using the comet assay. The intracellular levels of reactive oxygen species in oocytes was measured by 2 0 ,7 0-dichlorodihydrofluorescein diacetate fluorescence. The cryopreservation of ovarian tissue by the slow freezing technique resulted in a significantly higher level of DNA fragmentation in oocytes in comparison to vitrification (p50.05) whereas DNA fragmentation in granulosa cells was significantly higher than the control (p50.01). Further, reactive oxygen species were significantly elevated in oocytes derived from slow freezing when compared to vitrification (p50.05). Therefore, we conclude that the ovarian tissue slow freeze-thawing makes the oocyte and granulosa cells more vulnerable to DNA damage whereas vitrification appears to be a safer method than slow freezing for ovarian tissue cryopreservation.

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Section: Discussionmentioning

confidence: 99%

Ovarian tissue vitrification is more efficient than slow freezing in protecting oocyte and granulosa cell DNA integrity

Mathias

D’Souza

Uppangala

et al. 2014

Systems Biology in Reproductive Medicine

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“…Freezing forms an osmotic imbalance, which results in a flow of water from the inside of cells [8]. In this case, the concentration of ions and other solvents increases beyond physiological concentrations, which results in chemical stress [9][10][11][12]. With ice crystallization, the osmotic stress is increasing and such dehydration damages membranes and organelles [2].…”

Section: Introductionmentioning

confidence: 99%

Simple Syntheses of New Pegylated Trehalose Derivatives as a Chemical Tool for Potential Evaluation of Cryoprotectant Effects on Cell Membrane

et al. 2020

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In our work, we developed the synthesis of new polyfunctional pegylated trehalose derivatives and evaluated their cryoprotective effect using flow cytometry. We showed that new compounds (modified trehaloses) bound to appropriate extracellular polymeric cryoprotectants could be helpful as a chemical tool for the evaluation of their potential toxic cell membrane influences. Our aim was to form a chemical tool for the evaluation of cryoprotectant cell membrane influences, which are still not easily predicted during the freezing/thawing process. We combined two basic cryoprotectants: polyethyleneglycols (PEGs) and trehalose in the new chemical compounds—pegylated trehalose hybrids. If PEG and trehalose are chemically bound and trehalose is adsorbed on the cell surface PEGs molecules which are, due to the chemical bonding with trehalose, close to the cell surface, can remove the cell surface hydration layer which destabilizes the cell membrane. This was confirmed by the comparison of new material, PEG, trehalose, and their mixture cryoprotective capabilities.

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“…Many factors may affect the outcomes of embryo vitrification, such as the use of different CPAs (Berthelot-Ricou et al, 2013), embryo quality (Desai, Goldberg, Austin, & Falcone, 2013;Kattera & Chen, 2006;Seki, Jin, & Mazur, 2014;Seki & Mazur, 2012;Vanderzwalmen et al, 2002Vanderzwalmen et al, , 2009 and carriers (e.g. Cryotop), which influence the cooling and warming rates during vitrification and thawing (Kuwayama, 2007;Lane & Gardner, 2001;Martino, Songsasen, & Leibo, 1996;Nakashima et al, 2010;Seki & Mazur, 2012;Seki et al, 2014;Vanderzwalmen et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Shortened equilibration time can compromise clinical outcomes in human embryo vitrification

Xiong

Liu²,

Gao

et al. 2016

Human Fertility

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Vitrification is an important way to cryopreserve human embryos and the recommended time of embryo exposure to the vitrification solution is 1 min. However, practically speaking, the duration of embryos exposure to equilibration solution can vary from 5 to 15 min. The purpose of this study was to investigate the effect of different equilibration times on the outcomes of frozen-thawed embryo transfer cycles. The data were collected from our medical records from January 2012 to June 2013 and a total of 517 cycles were included. These cycles were divided into four groups according to the equilibration time: (i) 5-6 min; (ii) 7-8 min; (iii) 9-10 min and (iv) 11-12 min. The results show that there were no differences in terms of survival rate and fully intact embryo rate among the four groups. However, lower clinical pregnancy, embryo implantation and live birth rates were observed in the 5-6 min exposure group (54.6%, 31.9% and 48.2%, respectively) compared with the three other groups. The corresponding rates in the 9-10 min group (73.5%, 47.6% and 64.7%) were the highest. This study indicated that different equilibration times influenced the clinical outcomes of human embryo vitrification and vitrification with shortened equilibration time compromised the clinical outcomes. Appropriate prolongation of the equilibrium time would probably improve the clinical outcomes.

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Genotoxicity assessment of mouse oocytes by comet assay before vitrification and after warming with three vitrification protocols

Cited by 21 publications

References 34 publications

Ovarian tissue vitrification is more efficient than slow freezing in protecting oocyte and granulosa cell DNA integrity

Ovarian tissue vitrification is more efficient than slow freezing in protecting oocyte and granulosa cell DNA integrity

Simple Syntheses of New Pegylated Trehalose Derivatives as a Chemical Tool for Potential Evaluation of Cryoprotectant Effects on Cell Membrane

Shortened equilibration time can compromise clinical outcomes in human embryo vitrification

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