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The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR

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Abstract

Purpose

This study was conducted on the effects of vitrification cryotop method on gene expression of mature oocytes in Mus musculus.

Methods

Transcript analyses of three mouse genes, namely Mater, Hook1 and Sod1, were performed upon non-vitrified and vitrified oocytes with different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG),15%: 7.5% DMSO + 7.5% EG, and 30%: 15% DMSO + 15% EG, using cryotop following normalization of transcripts with Hprt1 by nested quantitative PCR.

Results

Vitrification caused down-regulation of Mater and Hook1 and up-regulation of Sod1 when lower concentrations of cryoprotectants were used as opposed to the control group. The relative expression of Sod1 in vit2 (30% v/v) was significantly higher than vit1 (15% v/v). Quantitative transcript analysis of Mater and Hook1 for the vit2 condition failed to produce any data. Survival rates were the same for both vitrification treatments and significantly lower than control group.

Conclusions

Although vit1 treatment had lower survival rate compared to control group, it demonstrated better stability comparing to vit2 based on the transcript analysis.

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Acknowledgments

Special thanks to Raquel Fialho in Portugal for her advice in the Lab. This work was supported by CITA-A, University of the Azores, Angra do Hero´ısmo, Portugal and Cellular and Molecular Biology Researcher Center (CMBRC), Medical School of Shaheed Beheshti University of Medical Sciences and Health Services, Tehran, Iran.

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Authors and Affiliations

  1. Cellular and Molecular Biology Research Center, Medical School of Shaheed Beheshti University of Medical Sciences and Health Services, Parvaneh St. Yaman St. Chamran highway, Tehran, Iran

    Afrooz Habibi & Ahmad Hosseini

  2. Faculty of Agriculture, Department of Agronomy and Plant Breeding, Shahrood University of Technology, Shahrood, Iran

    Naser Farrokhi

  3. Animal Reproduction, Department of Agrarian Sciences, University of the Azores, Angra do Heroísmo, Portugal

    Fernando Moreira da Silva

  4. Hospital Santo Espírito de Angra do Heroísmo, SEEBMO, Angra do Heroísmo, Azores, Portugal

    Bruno F. Bettencourt & Jácome Bruges-Armas

  5. Department of Anatomy, Medical School of Tehran University of Medical Sciences and Health Services, Tehran, Iran

    Fardin Amidi

Authors
  1. Afrooz Habibi
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  2. Naser Farrokhi
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  3. Fernando Moreira da Silva
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  4. Bruno F. Bettencourt
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  5. Jácome Bruges-Armas
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  6. Fardin Amidi
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  7. Ahmad Hosseini
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Corresponding author

Correspondence to Ahmad Hosseini.

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Capsule

This research investigated the effects of vitrification cryotop method on gene expression (Mater, Sod1 and Hook1) in metaphase II mouse oocytes by nested quantitative PCR

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Habibi, A., Farrokhi, N., Moreira da Silva, F. et al. The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR. J Assist Reprod Genet 27, 599–604 (2010). https://doi.org/10.1007/s10815-010-9453-0

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  • DOI: https://doi.org/10.1007/s10815-010-9453-0

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