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Preincubation with glutathione ethyl ester improves the developmental competence of vitrified mouse oocytes

  • Gamete Biology
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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Oocyte vitrification is currently used for human fertility preservation. However, vitrification damage is a problem caused by decreasing ooplasmic levels of glutathione (GSH). The GSH donor glutathione ethyl ester (GSH-OEt) can significantly increase the GSH content in oocytes. However, it is difficult to obtain oocyte from woman. To overcome this, we used mouse oocytes to replace human oocytes as a model of study.

Methods

Oocytes from B6D2F1 mice were preincubated for 30 min with 2.5 mmol/L GSH-OEt (GSH-OEt group), without GSH-OEt preincubation before vitrification (control vitrification group) or in nonvitrified oocytes (fresh group). After thawing, oocytes were fertilized for evaluating the developmental competence of embryos in vitro and in vivo. Immunofluorescence, Polscope equipment and quantitative reverse transcription polymerase chain reaction (RT–qPCR) were used to analyze damage, including mitochondrial distribution, reactive oxygen species (ROS) levels, spindle morphology, and gene expression levels (Bcl-2, BAX, and MnSOD).

Results

The rates of fertilization, 3–4 cell, blastocyst formation and expanded blastocysts were significantly higher (p < 0.05) in the GSH-OEt group (90.4%; 91.1%; 88.9% and 63.0%) than in the control (80.0%; 81.4%; 77.7% and 50.5%). Provided embryos overcame the 2-cell block and developed to the blastocyst stage, birth rates of all groups were similar. Vitrification altered mitochondrial distribution, increased ROS levels, and caused abnormal spindle morphology; GSH-OEt preincubation could improve such damage. RT–qPCR showed that the expression of Bcl-2 was lower in the control group compared with the GSH-OEt group; BAX and MnSoD expression levels were higher in the control group than in the GSH-OEt group (p < 0.05).

Conclusions

The beneficial effect of GSH-OEt preincubation occurred before the 2-cell stage.

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Funding

Natural Science Foundation of China (No. 81601342) from Dr Yijuan Sun

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  1. Zhichao Li and Ruihuan Gu contributed equally to this work.

Authors and Affiliations

  1. Shanghai Ji Ai Genetics & IVF Institute, Obstetrics & Gynecology Hospital, Fudan University, Shanghai, 200011, People’s Republic of China

    Zhichao Li, Ruihuan Gu & Yijuan Sun

  2. Reproductive Medical Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, People’s Republic of China

    Xiaowei Lu, Shen Zhao & Yun Feng

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  1. Zhichao Li
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  2. Ruihuan Gu
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  3. Xiaowei Lu
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  4. Shen Zhao
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  5. Yun Feng
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  6. Yijuan Sun
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Correspondence to Yun Feng or Yijuan Sun.

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Li, Z., Gu, R., Lu, X. et al. Preincubation with glutathione ethyl ester improves the developmental competence of vitrified mouse oocytes. J Assist Reprod Genet 35, 1169–1178 (2018). https://doi.org/10.1007/s10815-018-1215-4

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