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Allotransplantation of cryopreserved prepubertal mouse ovaries restored puberty and fertility without affecting methylation profile of Snrpn-DMR

  • Hong-Yan Wang
    Affiliations
    Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China

    Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China
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  • Yun-Hong Li
    Affiliations
    Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China
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  • Lei Sun
    Affiliations
    Research Center of Developmental Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai, People's Republic of China
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  • Xuan Gao
    Affiliations
    Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China
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  • Li You
    Affiliations
    Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China
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  • Yin Wang
    Affiliations
    Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China
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  • Jing-Long Ma
    Affiliations
    Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China
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  • Zi-Jiang Chen
    Correspondence
    Reprint requests: Professor Zi-Jiang Chen, Ph.D., Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, 324 Jingwu Road, Jinan, People's Republic of China 250021.
    Affiliations
    Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China
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Published:September 17, 2012DOI:https://doi.org/10.1016/j.fertnstert.2012.08.030

      Objective

      To evaluate the genetic safety of vitrification on the methylation imprints and the development and fertility potential of prepubertal mouse ovaries.

      Design

      Experimental animal study.

      Setting

      University-based fertility center.

      Animal(s)

      Institute of Cancer Research (ICR) 10-day-old female mice, 10-week-old adult female mice, and 12-week-old adult male mice.

      Intervention(s)

      Vitrification of juvenile mouse ovaries was performed using ED20 and EG5.5/30 solutions followed by retrieval of fresh and vitrified-warmed germinal vesicle (GV) oocytes for Snrpn differentially methylated regions (DMR) methylation analyses, collection of mature oocytes from superovulated ovarian grafts, in vitro fertility(IVF), and early embryonic development after heterotopic allotransplantation.

      Main Outcome Measure(s)

      Analysis of methylation status of Snrpn-DMR, percentage of fertilization, and blastocysts formation.

      Result(s)

      Methylation status of Snrpn-DMR from vitrified-warmed GV oocytes did not show significant alteration compared with that of controls, although a significant reduction of viable oocytes was observed. Puberty as well as endocrine function was restored, and no significant difference was shown in number of follicles, percentage of mice retaining fertility, and blastocyst formation among three groups.

      Conclusion(s)

      Our study proved that vitrification of prepubertal mouse ovaries did not alter the methylation profile of Snrpn-DMR and subsequent allotransplantation; IVF could restore the development and fertility potential.

      Key Words

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