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2010 Fiscal Year Final Research Report

Cryopreservation of oocytes and embryos by inducing the expression of cryoprotectant channels.

Research Project

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Project/Area Number 20380155
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied animal science
Research InstitutionKochi University

Principal Investigator

EDASHIGE Keisuke  Kochi University, 教育研究部・総合科学系, 教授 (30175228)

Co-Investigator(Kenkyū-buntansha) KASAI Magosaburo  高知大学, 教育研究部・総合科学系, 教授 (60152617)
KOSHIMOTO Chihiro  宮崎大学, フロンティア科学実験総合センター, 教授 (70295210)
Project Period (FY) 2008 – 2010
Keywords凍結保存 / 卵子 / 胚 / 水透過性 / 耐凍剤透過性 / アクアポリン
Research Abstract

We tried to develop a vitrification method by which mouse oocytes and embryos at various stages can be cryopreserved by a single method. First, we examined whether the membrane-permeability of mouse oocytes and embryos can be equalized, because the permeability largely affects the condition for vitrification. We tried to induce the expression of water/cryoprotectant channels (AQP) in oocytes to increase the low membrane-permeability. We also tried in morulae to decrease the permeability of AQP3 which is markedly expressed in morulae and blastocysts. The cryoprotectant-permeability of oocytes increased slightly by retinoic acid but the maturation rate decreased remarkably. It was reported that the permeability of AQP3 decreased markedly at low pH. However, the permeability to ethylene glycol of mouse morulae, which expressed AQP3 abundantly, increased at low pH. Therefore, we considered that it would be difficult to make the membrane-permeability of oocytes/embryos equal. Next, we tried to develop a new vitrification solution in which a high concentration of sucrose was included to reduce the effect of membrane-permeability of oocytes/embryos on the vitrification condition. With this solution, embryos from the 2-cell to morula stages were successfully vitrified in the same condition, and vitrified embryos could be transported with dry ice (at -79℃). The method would be useful for cryopreservation of mouse embryos at various stages.

  • Research Products

    (38 results)

All 2011 2010 2009 2008

All Journal Article (11 results) (of which Peer Reviewed: 11 results) Presentation (25 results) Book (2 results)

  • [Journal Article] Developmental ability of vitrified mouse oocytes expressing water channels.2011

    • Author(s)
      Yohei Yamaji, Shinsuke Seki, Kazutsugu Matsukawa, Chihiro Koshimoto, Magosaburo Kasai, Keisuke Edashige.
    • Journal Title

      The Journal of Reproduction and Development 57

      Pages: 403-108

    • Peer Reviewed
  • [Journal Article] Cryobiological properties of immature zebrafish oocytes assessed by their ability to be fertilized and develop into hatching embryos.2011

    • Author(s)
      Shinsuke Seki, Toshimitsu Kouya, Ryoma Tsuchiya, Delgado M.Valdez Jr., Bo Jin, Chihiro Koshimoto, Magosaburo Kasai, Keisuke Edashige.
    • Journal Title

      Cryobiology 62

      Pages: 8-14

    • Peer Reviewed
  • [Journal Article] Human C-reactive protein enhances thrombus formation after neointimal balloon injury in transgenic rabbits.2011

    • Author(s)
      S.Matsuda, A.Yamashita, Y.Sato, S.Kitajima, T.Koike, C.Sugita, S.Moriguchi-Goto, K.Hatakeyama, M.Takahashi, C.Koshimoto, Y. Matsuura, T.Iwakiri, Y.E. Chen, J.Fan, Y.Asada.
    • Journal Title

      The Journal of Thrombosis and Haemostasis 9

      Pages: 201-208

    • Peer Reviewed
  • [Journal Article] Equilibrium vitrification of mouse embryos.2010

    • Author(s)
      Bo Jin, Keiji Mochida, Atsuo Ogura, Eri Hotta, Yukiko Kobayashi, Kaori Ito, Go Egawa, Shinsuke Seki, Hiroshi Honda, Keisuke Edashige, Magosaburo Kasai.
    • Journal Title

      Biology of Reproduction 82

      Pages: 444-450

    • Peer Reviewed
  • [Journal Article] Effects of bile acids on rat hepatic microsomal type I 11β-hydroxyl steroid dehydrogenase.2010

    • Author(s)
      Yorio Maeda, Seiji Naganuma, Ichiro Niina, Akio Shinohara, Chihiro Koshimoto, Kazuhiro Kondo, Kazuo Chijiiwa.
    • Journal Title

      Steroids 75

      Pages: 164-168

    • Peer Reviewed
  • [Journal Article] Critical role of von Willebr and factorand platelet interaction in venous thromboembolism.2009

    • Author(s)
      Misaki Takahashi, Atsushi Yamashita, Sayaka Moriguchi-Goto, Kousuke Marutsuka, Yuichiro Sato, Hiroshi Yamamoto, Chihiro Koshimoto, Yujiro Asada.
    • Journal Title

      Histology and Histopathology 24

      Pages: 1391-1398

    • Peer Reviewed
  • [Journal Article] Maintenance of fertility in cryopreserved Indian gerbil (Tatera indica) spermatozoa.2009

    • Author(s)
      Chihiro Koshimoto, Daisuke Watanabe, Akio Shinohara, Tetsuo Morita.
    • Journal Title

      Cryobiology 58

      Pages: 303-307

    • Peer Reviewed
  • [Journal Article] Analysis of antibody response by temperature-sensitive measles vaccine strain in the cotton rat model.2009

    • Author(s)
      Takeshi Haga, Niho Murayama, Yuya Shimizu, Akatsuki Saito, Takumi Sakamoto, Tetsuo Morita, Katsuhiro Komase, Tetsuo Nakayama, Kazuyuki Uchida, Tetsuro Katayama, Akio Shinohara, Chihiro Koshimoto, Hiroshi Sato, Hironori Miyata, Kiyoaki Katahira, Yoshitaka Goto.
    • Journal Title

      Comparative Immunology, Microbiology and Infectious Diseases 32

      Pages: 395-406

    • Peer Reviewed
  • [Journal Article] The mechanism by which mouse spermatozoa are injured during freezing.2008

    • Author(s)
      Bo Jin, Chihiro Yamasaki, Naoko Yamada, Shinsuke Seki, Delgado M. Valdez Jr., Magosaburo Kasai, Keisuke Edashige.
    • Journal Title

      Journal of Reproduction and Development 54

      Pages: 265-269

    • Peer Reviewed
  • [Journal Article] Formation of extracellular and intracellular ice during warming of vitrified mouse morulae and its effect on embryo survival.2008

    • Author(s)
      Bo Jin, Kenji Kusanagi, Makiko Ueda, Shinsuke Seki, Delgado M. Valdez Jr., Keisuke Edashige, Magosaburo Kasai.
    • Journal Title

      Cryobiology 56

      Pages: 233-240

    • Peer Reviewed
  • [Journal Article] Development of a reliable in vitro maturation system for zebrafish oocytes.2008

    • Author(s)
      Shinsuke Seki, Toshimitsu Kouya, Ryoma Tsuchiya, Delgado M.Valdez Jr., Bo Jin, Takao Hara, Naoya Saida, Magosaburo Kasai, Keisuke Edashige.
    • Journal Title

      Reproduction 135

      Pages: 285-292

    • Peer Reviewed
  • [Presentation] Aquaporin 9 facilitates the movement of DMSO and acetamide through the plasma membrane in mouse morulae.2010

    • Author(s)
      Keisuke Edashige, Shinsuke Seki, Hayato Arimura, Kan Matsuo, Yu-ichi Nakata, Bo Jin, Kazutsugu Matsukawa, Chihiro Koshimoto, Magosaburo Kasai
    • Organizer
      第15回中国畜産獣医学会(動物繁殖分会)
    • Place of Presentation
      Tianjin, Ruby Hotel.
    • Year and Date
      20101015-20101018
  • [Presentation] マウスの卵子および胚への耐凍剤透過におけるAquaporin 9の役割2010

    • Author(s)
      有村隼, 中田裕一, 関信輔, 松尾完, 金波, 松川和嗣, 葛西孫三郎, 枝重圭祐
    • Organizer
      第103回日本繁殖生物学会大会
    • Place of Presentation
      十和田市,北里大学獣医学部
    • Year and Date
      20100902-20100904
  • [Presentation] 凍結乾燥したウシ体細胞による核移植胚作出の試み2010

    • Author(s)
      長尾さや子, 長瀬祐樹, 枝重圭祐, 葛西孫三郎, 保地眞一, 松川和嗣
    • Organizer
      第103回日本繁殖生物学会大会
    • Place of Presentation
      十和田市,北里大学獣医学部
    • Year and Date
      20100902-20100904
  • [Presentation] The role of aquaporin 9 in the movement of Me_2SO and acetamide in mouse morulae.2010

    • Author(s)
      Keisuke Edashige, Shinsuke Seki, Hayato Arimura, Kan Matsuo, Yu-ichi Nakata, Bo Jin, Chihiro Koshimoto, Magosaburo Kasai
    • Organizer
      47th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Bristol, Bristol Royal Marriott Hotel.
    • Year and Date
      20100717-20100720
  • [Presentation] Handy transportation of vitrified mouse embryos with dry ice.2010

    • Author(s)
      Bo Jin, Keiji Mochida, Atsuo Ogura, Magosaburo Kasai, Keisuke Edashige
    • Organizer
      47th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Bristol, Bristol Royal Marriott Hotel.
    • Year and Date
      20100717-20100720
  • [Presentation] 耐凍剤チャンネル発現の誘導によるマウス卵子の耐凍性向上の試み2010

    • Author(s)
      和田紗希子, 中田裕一, 金波, 越本知大, 葛西孫三郎, 枝重圭祐
    • Organizer
      第57回日本実験動物学会総会
    • Place of Presentation
      京都,京都テルサ
    • Year and Date
      20100512-20100514
  • [Presentation] LASセミナー3:胚・精子の凍結保存2010

    • Author(s)
      中潟直己, 枝重圭祐
    • Organizer
      第57回日本実験動物学会総会
    • Place of Presentation
      京都,京都テルサ
    • Year and Date
      20100512-20100514
  • [Presentation] マウス卵巣のガラス化凍結保存2009

    • Author(s)
      金波, 本田宙, 西門優, 葛西孫三郎, 枝重圭祐
    • Organizer
      第102回日本繁殖生物学会大会
    • Place of Presentation
      奈良,近畿大学農学部
    • Year and Date
      20090910-20090912
  • [Presentation] Effect of the expression of aquaporins 1 and 3 in mouse MII oocytes on the nucleation temperature for intracellular ice formation.2009

    • Author(s)
      Shinsuke Seki, Keisuke Edashige, Peter Mazur
    • Organizer
      46th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Sapporo, Conference Hall on Hokkaido University.
    • Year and Date
      20090719-20090723
  • [Presentation] Rapid movement of water and cryoprotectants in pig expanded blastocysts via channel processes -its relevance to their tolerance to cryopreservation-.2009

    • Author(s)
      Keisuke Edashige, Bo Jin, Ryu-ichi Higashiyama, Jun-ichi Yonezawa, Masashi Miyake, Sei-ichi Takahashi, Ken-ichi Yazawa, Magosaburo Kasai
    • Organizer
      46th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Sapporo, Conference Hall on Hokkaido University.
    • Year and Date
      20090719-20090723
  • [Presentation] Equilibrium vitrification of mouse embryos.2009

    • Author(s)
      Bo Jin, Eri Hotta, Yukiko Kobayashi, Kaori Ito, Go Egawa, Shinsuke Seki, Hiroshi Honda, Keiji Mochida, Atsuo Ogura, Keisuke Edashige, Magosaburo Kasai
    • Organizer
      46th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Sapporo, Conference Hall on Hokkaido University.
    • Year and Date
      20090719-20090723
  • [Presentation] Survival of oocytes in antral follicles of vitrified mouse ovaries.2009

    • Author(s)
      Keisuke Edashige, Delgado M. Valdez Jr., Hiroshi Honda, Yu Nishikado, Bo Jin, Magosaburo Kasai
    • Organizer
      46th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Sapporo, Conference Hall on Hokkaido University.
    • Year and Date
      20090719-20090723
  • [Presentation] ドライシッパーを用いないマウス胚の冷蔵及びドライアイス温度下での輸送の検討2009

    • Author(s)
      持田慶司, 鬼頭靖司, 金波, 枝重圭祐, 葛西孫三郎, 小倉淳郎
    • Organizer
      第56回日本実験動物学会総会
    • Place of Presentation
      大宮,大宮ソニックシティ
    • Year and Date
      20090514-20090516
  • [Presentation] 様々な発育ステージのガラス化凍結マウス胚の-80℃での短期保持-ドライアイスを用いた簡便な輸送を目指して-2009

    • Author(s)
      金波, 堀田英里, 小林由希子, 伊藤香, 関信輔, 本田宙, 持田慶司, 小倉淳郎, 葛西孫三郎, 枝重圭祐
    • Organizer
      第56回日本実験動物学会総会
    • Place of Presentation
      大宮,大宮ソニックシティ
    • Year and Date
      20090514-20090516
  • [Presentation] LASセミナー3:胚・精子の凍結保存2009

    • Author(s)
      中潟直己, 枝重圭祐
    • Organizer
      第56回日本実験動物学会総会
    • Place of Presentation
      大宮,大宮ソニックシティ
    • Year and Date
      20090514-20090516
  • [Presentation] ブタの卵子と拡大胚盤胞における水と耐凍剤の透過経路2009

    • Author(s)
      金波, 東山龍一, 米澤潤一, 高橋誠一, 三宅正史, 葛西孫三郎, 枝重圭祐
    • Organizer
      第110回日本畜産学会大会
    • Place of Presentation
      藤沢市,日本大学生物資源科学部
    • Year and Date
      20090327-20090929
  • [Presentation] Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio).2008

    • Author(s)
      Shinsuke Seki, Bo Jin, Tsuchiya Ryouma, Toshimitsu Kouya, Takao Hara, Delgado M.Valdez Jr., Magosaburo Kasai, Keisuke Edashige.
    • Organizer
      The 5th Annual Conference of the Asia Reproductive Biotechnology Society
    • Place of Presentation
      Kunming, Horizon Hotel.
    • Year and Date
      20081127-20081201
  • [Presentation] Short-term storage of vitrified two-cell mouse embryos at -80oC for handy transportation with dry ice.2008

    • Author(s)
      Bo Jin, Eri Hotta, Yukiko Kobayashi, Shinsuke Seki, Masahiro Yoshimura, Hiroshi Honda, Keiji Mochida, Atsuo Ogura, Magosaburo Kasai, Keisuke Edashige
    • Organizer
      第101回日本繁殖生物学会大会
    • Place of Presentation
      福岡市,九州大学医学部百年講堂
    • Year and Date
      20080918-20080920
  • [Presentation] マウス桑実胚の耐凍剤透過性におけるaquaporin 9の役割2008

    • Author(s)
      松尾完, 関信輔, 金波, Valdez M.Delgado Jr., 太田悟史, 好村正博, 葛西孫三郎, 枝重圭祐
    • Organizer
      第101回日本繁殖生物学会大会
    • Place of Presentation
      福岡市,九州大学医学部百年講堂
    • Year and Date
      20080918-20080920
  • [Presentation] マウス精子が凍結保存によって受ける傷害のメカニズム2008

    • Author(s)
      好村正博, 金波, 山崎千寛, 山田奈央子, 関信輔, Valdez M.Delgado Jr., 葛西孫三郎, 枝重圭祐
    • Organizer
      第101回日本繁殖生物学会大会
    • Place of Presentation
      福岡市,九州大学医学部百年講堂
    • Year and Date
      20080918-20080920
  • [Presentation] ウシ桑実胚の水透過と耐凍剤透過におけるaquaporin 3の役割2008

    • Author(s)
      河合泰典, 原隆夫, 松尾完, Valdez M.Delgado Jr., 金波, 好村正博, 東山龍一, 市川恭子, 恒石望太郎, 枝重圭祐, 葛西孫三郎
    • Organizer
      第101回日本繁殖生物学会大会
    • Place of Presentation
      福岡市,九州大学医学部百年講堂
    • Year and Date
      20080918-20080920
  • [Presentation] Effects of raffinose concentration and methyl-beta cyclodextrin on IVF with cryopreserved C57BL/6 sperm.2008

    • Author(s)
      Keiji Mochida, Chihiro Koshimoto, Keisuke Edashige, Kentaro Aizawa, Kyuichi Taguma, Akihiko Ohta, Atsuo Ogura
    • Organizer
      45th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Charlotte, The Omni Charlotte Hotel.
    • Year and Date
      20080720-20080723
  • [Presentation] The pathway for the movement of water and cryoprotectants in bovine oocytes and embryos.2008

    • Author(s)
      Keisuke Edashige, Takao Hara, Yasunori Kawai, Masahiro Yoshimura, Bo Jin, Delgado M.Valdez, Jr., Magosaburo Kasai
    • Organizer
      45th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Charlotte, The Omni Charlotte Hotel.
    • Year and Date
      20080720-20080723
  • [Presentation] Permeability to water and cryoprotectants of zebrafish (Danio Rerio) oocytes at mid stage III.2008

    • Author(s)
      Delgado M.Valdez Jr., Ryoma Tsuchiya, Shinsuke Seki, Naoya Saida, Bo Jin, Masahiro Yoshimura, Chihiro Koshimoto, Magosaburo Kasai, Keisuke Edashige
    • Organizer
      45th Annual Meeting of the Society for Cryobiology
    • Place of Presentation
      Charlotte, The Omni Charlotte Hotel.
    • Year and Date
      20080720-20080723
  • [Presentation] 動物細胞と凍結-哺乳動物の卵子と受精卵の耐凍性に関わるタンパク質-2008

    • Author(s)
      枝重圭祐
    • Organizer
      第54回日本生物工学会大会セミナー
    • Place of Presentation
      石川郡野々市町,石川県立大学
    • Year and Date
      20080613-20080614
  • [Book] Fertility Cryopreservation(Movement of water and cryoprotectants in mouse oocytes and embryos at different stages : relevance to cryopreservation)(Ri-Cheng Chian, Patrick Quinn (Eds.))2010

    • Author(s)
      葛西孫三郎, 枝重圭祐
    • Total Pages
      16-23
    • Publisher
      Cambridge University Press
  • [Book] 生殖系列細胞の保存(生命の誕生に向けて(第二版)生殖補助医療(ART)培養の理論と実際(日本哺乳動物卵子学会編)2010

    • Author(s)
      葛西孫三郎, 枝重圭祐
    • Total Pages
      27-34
    • Publisher
      近代出版

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Published: 2012-01-26   Modified: 2016-04-21  

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