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Establishment of an efficient cryopreservation method for immature porcine oocytes

Research Project

Project/Area Number 26870839
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Conservation of biological resources
Animal production science
Research InstitutionNational Agriculture and Food Research Organization

Principal Investigator

Somfai Tamas  国立研究開発法人農業・食品産業技術総合研究機構, 畜産研究部門・家畜育種繁殖研究領域, 上級研究員 (90547720)

Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsブタ / ジーンバンク / 生殖補助医療技術 / 卵子 / 超低温保存 / ガラス化保存 / 発生能 / vitrification / porcine / immature oocyte / gene expression / cytoplasmic maturation / apoptosis / maturation medium / embryo development / 未成熟卵子 / レドックス状態 / 成熟培地 / 胚発生 / アポトーシス / 遺伝発現 / 遺伝子発現
Outline of Final Research Achievements

To improve the efficacy for embryo production from cryopreserved porcine oocytes, we investigated cytoplasmic damages caused by vitrification and optimized the vitrification and subsequent culture protocol. Vitrification did not affect dramatically the cytoplasmic maturation, redox status, energy levels and RNA content in oocytes and gap junctional communication between oocytes and cumulus cells but caused premature nuclear resumption and reversible damage in nucleolus and microfilaments in oocytes. We optimized the vitrification procedure by defining the best cryoprotectant combination, treatment regimen and temperature. Supplementation of the culture medium with the antiapoptotic agent resveratrol during post-warming maturation culture improved the ability of vitrified oocytes to develop to the blastocyst stage. By these modifications we could set up a simplified defined cryopreservation system for porcine oocytes with improved efficacy according to the aim of the project.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (18 results)

All 2017 2016 2015 2014

All Journal Article (3 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 2 results,  Acknowledgement Compliant: 2 results) Presentation (14 results) (of which Int'l Joint Research: 4 results,  Invited: 3 results) Book (1 results)

  • [Journal Article] Effects of polyethylene glycol and a synthetic ice blocker during vitrification of immature porcine oocytes on survival and subsequent embryo development2017

    • Author(s)
      1.Santos EC, Somfai T, Appeltant R, Dang-Nguyen TQ, Noguchi J, Kaneko H, Kikuchi K
    • Journal Title

      Animal Science Journal

      Volume: in press Issue: 8 Pages: 1042-1048

    • DOI

      10.1111/asj.12730

    • NAID

      120006473162

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] The effects of vitrification at the germinal vesicle stage of cumulus enclosed porcine oocytes on cumulus expansion, nuclear progression and cytoplasmic maturation2017

    • Author(s)
      2.Appeltant R, Somfai T, Santos E, Dang-Nguyen TQ, Nagai T, Kikuchi K
    • Journal Title

      Reproduction, Fertility and Development

      Volume: in press

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Optimization of cryoprotectant treatment for the vitrification of immature cumulus-enclosed porcine oocytes: comparison of sugars, combinations of permeating cryoprotectants and equilibration regimens2015

    • Author(s)
      Somfai T, Men NT, Noguchi J, Kaneko H, Kashiwazaki N, Kikuchi K.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 61 Issue: 6 Pages: 571-579

    • DOI

      10.1262/jrd.2015-089

    • NAID

      130005113568

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2015 Research-status Report
  • [Presentation] The effect of exposure time on the toxicity of vitrification solution on porcine cumulus-oocyte complexes before in vitro maturation2017

    • Author(s)
      Appeltant R, Somfai T, Santos ECS, Kikuchi K
    • Organizer
      The 43th Annual Meeting of the International Embryo Transfer Society
    • Place of Presentation
      Austin (アメリカ合衆国)
    • Year and Date
      2017-01-14
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The effects of resveratrol during in vitro maturation on the developmental competence of porcine oocytes vitrified at the immature stage2017

    • Author(s)
      4.Santos ECS, Somfai T, Appeltant R, Dang-Nguyen TQ, Kaneko H, Noguchi J, Nagai T, Kikuchi K
    • Organizer
      The 43th Annual Meeting of the International Embryo Transfer Society
    • Place of Presentation
      Austin (アメリカ合衆国)
    • Year and Date
      2017-01-14
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Oocyte cryopreservation for gene banking in pigs2016

    • Author(s)
      Somfai T.
    • Organizer
      The 17th AAAP Animal Science Congress (Symposium 07- Current Status of Animal Biotechnology and the Application for
    • Place of Presentation
      九州産業大学(福岡県・福岡)
    • Year and Date
      2016-08-22
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] The effects of vitrification at the germinal vesicle stage on transzonal projections and cumulus expansion in porcine cumulus-oocyte complexes.2016

    • Author(s)
      Appeltant R, Somfai T, Kikuchi K.
    • Organizer
      18th International Congress on Animal Reproduction (ICAR)
    • Place of Presentation
      Tours (フランス)
    • Year and Date
      2016-06-26
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The effects of polyethylene glycol and a synthetic ice blocker on survival and development of immature porcine oocytes vitrified by the Cryotop method.2016

    • Author(s)
      6.Santos EC, Somfai T, Appeltant R, Dang-Nguyen TQ, Kikuchi K
    • Organizer
      18th International Congress on Animal Reproduction (ICAR)
    • Place of Presentation
      Tours (フランス)
    • Year and Date
      2016-06-26
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The effects of polyethylene glycol and Supercool X-1000 during the vitrification of immature porcine oocytes.2016

    • Author(s)
      Santos ECD, Somfai T, Kikuchi K
    • Organizer
      第121回日本畜産学会
    • Place of Presentation
      日本獣医生命科学大学
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Presentation] Vitrification at the germinal vesicle stage triggers precocious meiotic resumption but does not affect cytoplasmic maturation in cumulus-enclosed porcine oocytes during in vitro maturation.2016

    • Author(s)
      Somfai T, Men NT, Kaneko H, Noguchi J, Haraguchi S, Santos ECD, Nagai T, Kikuchi K.
    • Organizer
      The 42th Annual Conference of the International Embryo Transfer Society
    • Place of Presentation
      Louisville, Kentucky, USA
    • Year and Date
      2016-01-23
    • Related Report
      2015 Research-status Report
  • [Presentation] Update on the cryopreservation of porcine oocytes2015

    • Author(s)
      Somfai T, Kikuchi K, Kaneko H, Noguchi J, Men NT, Santos ECD, Nagai T.
    • Organizer
      3rd Fatty Pig Science and Utilization International Conference
    • Place of Presentation
      Herceghalom, Hungary
    • Year and Date
      2015-11-18
    • Related Report
      2015 Research-status Report
  • [Presentation] Comparison of sugars, combinations of permeable cryoprotectants, and equilibration regimens for the solid surface vitrification of immature porcine oocytes.2015

    • Author(s)
      Somfai Tamas, Men NT., Kaneko H., Noguchi J., 原口 清輝, 永井 卓, 菊地 和弘
    • Organizer
      The 41th Annual Conference of the International Embryo Transfer Society
    • Place of Presentation
      Versailles(France)
    • Year and Date
      2015-01-13
    • Related Report
      2014 Research-status Report
  • [Presentation] The effects of collection season and storage duration in liquid nitrogen on post-warming survival and nuclear maturation of immature porcine oocytes preserved by solid surface vitrification.2015

    • Author(s)
      永井 卓, Somfai Tamas, Nguyen Thi Men, 金子 浩之, 谷原 史倫, 菊地 和弘
    • Organizer
      The 41th Annual Conference of the International Embryo Transfer Society
    • Place of Presentation
      Versailles(France)
    • Year and Date
      2015-01-12
    • Related Report
      2014 Research-status Report
  • [Presentation] Recent progress in the vitrification of porcine oocytes.2014

    • Author(s)
      Somfai Tamas, 吉岡 耕治, 柏崎 直巳, Istvan Egerszegi, Jozsef Ratky, 永井 卓, 菊地 和弘
    • Organizer
      The 11th Annual Conference of the Asian Reproductive Biotechnology Society
    • Place of Presentation
      Bangkok(Thailand)
    • Year and Date
      2014-11-03
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Recent progress in the cryopreservation of immature porcine oocytes.2014

    • Author(s)
      Somfai Tamas, 菊地 和弘, 吉岡 耕治, 柏崎 直巳, 永井 卓
    • Organizer
      第107回日本繁殖生物学会大会
    • Place of Presentation
      帯広畜産大学(北海道帯広市)
    • Year and Date
      2014-08-24
    • Related Report
      2014 Research-status Report
  • [Presentation] The effects of season, medium storage at 4 C and sample storage in liquid nitrogen on post-warming survival of vitrified porcine oocytes.2014

    • Author(s)
      Somfai Tamas, Istvan Egerszegi, Nguyen Thi Men, 金子 浩之, 野口 純子, Jozsef Ratky,柏崎 直巳, 菊地 和弘
    • Organizer
      第107回日本繁殖生物学会大会
    • Place of Presentation
      帯広畜産大学(北海道帯広市)
    • Year and Date
      2014-08-21
    • Related Report
      2014 Research-status Report
  • [Presentation] Cryopreservation of in vitro produced embryos and immature oocytes in pigs.2014

    • Author(s)
      Somfai Tamas, 菊地 和弘, 吉岡 耕治, 柏崎 直巳, 永井 卓
    • Organizer
      International Symposium on Cutting-Edge Reproductive Technologies and Perspectives for their Usage in Swine
    • Place of Presentation
      Tainan(Taiwan)
    • Year and Date
      2014-06-05
    • Related Report
      2014 Research-status Report
    • Invited
  • [Book] Synchronization of In Vitro Maturation in Porcine Oocytes ( in "Cell Cycle Synchronization", 2nd edition, Ed: Banfalvi G.)2017

    • Author(s)
      Somfai T, Hirao Y.
    • Publisher
      Humana Press (Springer)
    • Related Report
      2016 Annual Research Report

URL: 

Published: 2014-04-04   Modified: 2018-03-22  

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