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Improved technology of mouse somatic cell nuclear transfer that greatly increases cloning efficiency

Research Project

Project/Area Number 23580390
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Applied animal science
Research InstitutionKyoto University

Principal Investigator

YAMADA Masayasu  京都大学, (連合)農学研究科(研究院), 准教授 (10243073)

Project Period (FY) 2011 – 2013
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Keywords体細胞核移植 / クローンマウス / ヒストン脱アセチル化酵素阻害剤 / 脱イオン化BSA / ビタミンC / エピジェネティックス / X染色体不活化異常 / 脱イオン化BSA注入 / trichostatin A / vitamin C / Oct3/4の発現 / ヒストンアセチル化 / ヒストンメチル化 / マウス / 胚盤胞 / Oct4発現 / H3K27me3発現 / 着床後発生 / 着床前発生 / エピジェネティックス調節
Research Abstract

Production rate of cloned mice still is very low, which is thought to be attributed to abnormal epigenetic modifications and X-chromosomal inactivation (XCI) status in somatic cell nuclear transfer (SCNT) embryos. Here, we developed novel mouse SCNT technologies as follows; 1) treatment with valproi acid for 24 h from the 4-cell stage of SCNT embryos, which improved expression of Oct4 and trimethylated histone H3 lysine 27, a marker of the XCI status in SCNT blastocysts, 2) intracytoplasmic injection of deionized BSA after SCNT, which promoted expression of acetylated histone H3 lysine 9 and acetylated histone H4 lysine 12 at the pronuclear stage and production of cloned offspring, and 3) treatments with tricostatine A for 8 h from the beginning of activation of SCNT oocytes followed by vitamin C for 7 h, which promoted the oxidation of 5-methylcytocine to 5-hydroxymethylcytosine in pronuclear DNA of SCNT embryos and led to greatly increased production of cloned offspring.

Report

(4 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • 2011 Research-status Report
  • Research Products

    (37 results)

All 2014 2013 2012 2011 Other

All Journal Article (14 results) (of which Peer Reviewed: 10 results) Presentation (20 results) Book (2 results) Remarks (1 results)

  • [Journal Article] Midkine and cytoplasmic maturation of mammalian oocytes in the context of ovarian follicle physiology.2014

    • Author(s)
      Ikeda S, Yamada M
    • Journal Title

      BRITISH JOURNAL OF PHARMACOLOGY

      Volume: 171 Issue: 4 Pages: 827-836

    • DOI

      10.1111/bph.12311

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Valproic acid treatment from the 4-cell stage improves Oct4 expression and nuclear distribution of histone H3K27me3 in mouse cloned blastocysts2013

    • Author(s)
      Isaji Y, Murata M, Takaguchi N, Mukai T, Tajima Y, Imai H, Yamada M.
    • Journal Title

      J. Reprod. Dev

      Volume: 59(2) Pages: 196-204

    • NAID

      10031170779

    • Related Report
      2013 Final Research Report
  • [Journal Article] Valproic Acid Treatment from the 4-cell Stage Improves Oct4 Expression and Nuclear Distribution of Histone H3K27me3 in Mouse Cloned Blastocysts2013

    • Author(s)
      Isaji Y, Murata M, Takaguchi N, Mukai T, Tajima Y, Imai H, Yamada M
    • Journal Title

      Journal of Reproduction and Development

      Volume: 59 Issue: 2 Pages: 196-204

    • DOI

      10.1262/jrd.2012-156

    • NAID

      10031170779

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Effects of extracellular matrices and lectin Dolichos biflorus agglutinin on cell adhesion and self-renewal of bovine gonocytes cultured in vitro2013

    • Author(s)
      Kim SM, Fujihara M, Sahare M, Minami N, Yamada M and Imai H
    • Journal Title

      Reproduction Fertility and Development

      Volume: In press Issue: 2 Pages: 268-268

    • DOI

      10.1071/rd12214

    • NAID

      120005367805

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Valproic Acid Treatment from the 4-cell Stages Improves Oct4 Expression and Nuclear Distribution of Histone H3K27me3 in Mouse Cloned Blastocysts.2013

    • Author(s)
      Isaji Y, Murata M, Takaguchi N, Mukai T, Tajima Y, Imai H, Yamada M
    • Journal Title

      J Reprod Dev

      Volume: in press

    • NAID

      10031170779

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] ノコダゾール処理除核卵母細胞由来マウス体細胞核移植胚の発生に及ぼすスクリプタイドの効果2012

    • Author(s)
      伊佐治優希、村田萌子、田島陽介、今井裕、山田雅保
    • Journal Title

      日本胚移植学雑誌

      Volume: 34 Pages: 117-127

    • NAID

      10031130927

    • Related Report
      2013 Final Research Report
  • [Journal Article] ノコダゾール処理除核卵母細胞由来マウス体細胞核移植の発生に及ぼすスクリプタイドの効果2012

    • Author(s)
      伊佐治優希、村田萌子、田島陽介、今井裕、山田雅保
    • Journal Title

      日本胚移植学雑誌

      Volume: 34 Pages: 117-127

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system2011

    • Author(s)
      Tsukiyama T, Asano R, Kawaguchi T, Kim N, Yamada M, Minami N, Ohinata Y, Imai H.
    • Journal Title

      Genes Cells

      Volume: 16(7) Pages: 815-825

    • NAID

      120005593680

    • Related Report
      2013 Final Research Report
  • [Journal Article] Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos2011

    • Author(s)
      Miyamoto K, Nagai K, Kitamura N, Nishikawa T, Ikegami H, Binh NT, Tsukamoto S, Matsumoto M, Tsukiyama T, Minami N, Yamada M, Ariga H, Miyake M, Kawarasaki T, Matsumoto K, Imai H.
    • Journal Title

      Proc. Natl. Acad. Sci. USA

      Volume: 108(17) Pages: 7040-7045

    • Related Report
      2013 Final Research Report
  • [Journal Article] Characterization and in vitro culture of male germ cells from developing bovine testis.2011

    • Author(s)
      Fujihara M, Kim SM, Minami N, Yamada M, Imai H
    • Journal Title

      J Reprod Dev

      Volume: 57 Pages: 355-364

    • NAID

      10029050501

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos.2011

    • Author(s)
      Miyamoto K, Nagai K, Kitamura N, Nishikawa T, Ikegami H, Binh NT, Tsukamoto S, Matsumoto M, Tsukiyama T, Minami N, Yamada M, Ariga H, Miyake M, Kawarasaki T, Matsumoto K, Imai
    • Journal Title

      Proc Natl Acad Sci U S A

      Volume: 108 Pages: 7040-7045

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system.2011

    • Author(s)
      Tsukiyama T, Asano R, Kawaguchi T, Kim N, Yamada M, Minami N, Ohinata Y, Imai H
    • Journal Title

      Genes Cells

      Volume: 16 Pages: 815-825

    • NAID

      120005593680

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Review: Structural and functional changes linked to, and factors promoting, cytoplasmic maturation in mammalian oocytes.2011

    • Author(s)
      Yamada M, Isaji Y
    • Journal Title

      Reprod Med Biol

      Volume: 10 Pages: 69-79

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] 卵子成熟と遺伝子発現2011

    • Author(s)
      山田雅保
    • Journal Title

      Hormone Frontier in Gynecology

      Volume: 18 Pages: 27-31

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Presentation] マウス体細胞クローン胚の発生に及ぼすビタミンCの効果2013

    • Author(s)
      田島陽介、伊佐治優希、後藤奈々、今井裕、山田雅保
    • Organizer
      第106回日本繁殖生物学会大会
    • Place of Presentation
      府中
    • Year and Date
      2013-09-12
    • Related Report
      2013 Final Research Report
  • [Presentation] マウス体細胞クローン胚の発生に及ぼすビタミンCの効果2013

    • Author(s)
      田島陽介、伊佐治優希、後藤奈々、今井裕、山田雅保
    • Organizer
      第106回日本繁殖生物学会
    • Place of Presentation
      東京農工大学(東京都府中市)
    • Related Report
      2013 Annual Research Report
  • [Presentation] ウシにおけるマウスES 細胞様iPS 細胞の樹立の試み2013

    • Author(s)
      川口高正、南直治郎、山田雅保、今井 裕、木村康二
    • Organizer
      第1回 関西生殖医学集談会、第45回 関西アンドロロジーカンファレンス
    • Place of Presentation
      ハービスPLAZA会議室(大阪市)
    • Related Report
      2012 Research-status Report
  • [Presentation] Generation of murine-ES-cell-like iPS cells in cattle using piggyBac transposition of doxycycline-inducible transcription factor2013

    • Author(s)
      Kawaguchi T, Tsukiyama T, Minami N, Yamada M, Kimura K, Imai H
    • Organizer
      CiRA International Symposium 2013 ~Raising the next generation of stem cell research~
    • Place of Presentation
      Kyoto University, Kyoto
    • Related Report
      2012 Research-status Report
  • [Presentation] マウス体細胞核移植胚の着床前発生に及ぼすビタミンCの効果2012

    • Author(s)
      田島陽介、伊佐治優希、今井裕、山田雅保
    • Organizer
      第105回日本繁殖生物学会大会
    • Place of Presentation
      つくば
    • Year and Date
      2012-09-08
    • Related Report
      2013 Final Research Report
  • [Presentation] 脱イオン化ウシ血清アルブミンのマウス体細胞核移植胚への注入はその胚盤胞への発生およびヒストンアセチル化を促進する2012

    • Author(s)
      伊佐治優希、田島陽介、今井裕、山田雅保
    • Organizer
      第105回日本繁殖生物学会大会
    • Place of Presentation
      つくば
    • Year and Date
      2012-09-08
    • Related Report
      2013 Final Research Report
  • [Presentation] 脱イオン化ウシ血清アルブミンのマウス体細胞核移植胚への注入はその胚盤胞への発生を促進する2012

    • Author(s)
      伊佐治優希、西川智章、今井裕、山田雅保
    • Organizer
      日本畜産学会第115回大会
    • Place of Presentation
      名古屋
    • Year and Date
      2012-03-28
    • Related Report
      2013 Final Research Report
  • [Presentation] 脱イオン化ウシ血清アルブミンのマウス体細胞核移植胚への注入は、その胚盤胞期への発生およびヒストンアセチル化を促進する2012

    • Author(s)
      伊佐治優希、田島陽介、今井裕、山田雅保
    • Organizer
      第105回日本繁殖生物学会大会
    • Place of Presentation
      筑波大学大学会館
    • Related Report
      2012 Research-status Report
  • [Presentation] マウス体細胞核移植胚の着床前発生に及ぼすビタミンCの効果2012

    • Author(s)
      田島陽介、伊佐治優希、今井裕、山田雅保
    • Organizer
      第105回日本繁殖生物学会大会
    • Place of Presentation
      筑波大学大学会館
    • Related Report
      2012 Research-status Report
  • [Presentation] Derivation of murine-ES-cell like iPS cells in canine under the EOS reporter system2012

    • Author(s)
      Kawaguchi T, Asano R, Tsukiyama T, Minami N, Yamada M, Hatoya S, Sugiura K, Inaba T, Imai H
    • Organizer
      International Society for Stem Cell Research (ISSCR)
    • Place of Presentation
      Pacifico Yokohama
    • Related Report
      2012 Research-status Report
  • [Presentation] Transdifferentiation of bovine aminion cells into trophoblast-like cells using piggyBac transposition of doxycycline-inducible reprogramming factors2012

    • Author(s)
      Kawaguchi T, Tsukiyama T, Minami N, Yamada M, Kimura K, Imai H
    • Organizer
      Satellite symposium to the International Congress on Animal Reproduction (ICAR)
    • Place of Presentation
      Banff, Canada
    • Related Report
      2012 Research-status Report
  • [Presentation] 活性化後のバルプロ酸処理がマウス体細胞核移植胚の胚盤胞への発生およびOct4の発現に及ぼす影響2011

    • Author(s)
      伊佐治優希、村田萌子、高口尚也、今井裕、山田雅保
    • Organizer
      第104回日本繁殖生物学会大会
    • Place of Presentation
      盛岡
    • Year and Date
      2011-09-15
    • Related Report
      2013 Final Research Report
  • [Presentation] マウス体細胞核移植における化学的処理による除核方法の簡便化についての検討2011

    • Author(s)
      村田萌子、伊佐治優希、田島陽介、今井裕、山田雅保
    • Organizer
      第61回関西畜産学会大会
    • Place of Presentation
      岡山
    • Year and Date
      2011-09-13
    • Related Report
      2013 Final Research Report
  • [Presentation] マウス体細胞核移植における化学的処理による除核条件の検討2011

    • Author(s)
      村田萌子、伊佐治優希、田島陽介、今井裕、山田雅保
    • Organizer
      第18回日本胚移植研究会大会
    • Place of Presentation
      神戸
    • Year and Date
      2011-09-08
    • Related Report
      2013 Final Research Report
  • [Presentation] 活性化後のバルプロ酸処理がマウス体細胞核移植胚の胚盤胞期への発生およびOct4の発現に及ぼす影響

    • Author(s)
      伊佐治優希、村田萌子、高口尚也、今井裕、山田雅保
    • Organizer
      第104回日本繁殖生物学会大会
    • Place of Presentation
      いわて県民情報交流センター・アイーナ
    • Related Report
      2011 Research-status Report
  • [Presentation] ウシ血清アルブミン中の混入物質が体外培養におけるマウス胚盤胞の長期的形態及び機能維持に及ぼす効果

    • Author(s)
      高口尚也、向井利太、伊佐治優希、今井裕、山田雅保
    • Organizer
      第104回日本繁殖生物学会大会
    • Place of Presentation
      いわて県民情報交流センター・アイーナ
    • Related Report
      2011 Research-status Report
  • [Presentation] マウス体細胞核移植における化学的処理による除核方法の簡便化についての検討

    • Author(s)
      村田萌子、伊佐治優希、田島陽介、今井裕、山田雅保
    • Organizer
      第61回関西畜産学会大会
    • Place of Presentation
      岡山大学創立五十周年記念館
    • Related Report
      2011 Research-status Report
  • [Presentation] 体外培養におけるマウス胚盤胞の形態及び機能の長期的維持に関する研究 ー脱イオン化処理BSAの効果ー

    • Author(s)
      向井 利太, 高口 尚也, 伊佐治 優希, 今井 裕, 山田 雅保
    • Organizer
      第18回日本胚移植研究会兵庫大会
    • Place of Presentation
      神戸大学百年記念館(神大会館)六甲ホール
    • Related Report
      2011 Research-status Report
  • [Presentation] マウス体細胞核移植における化学的処理による除核条件の検討

    • Author(s)
      村田萌子・伊佐治優希・田島陽介・今井裕・山田雅保
    • Organizer
      第18回日本胚移植研究会兵庫大会
    • Place of Presentation
      神戸大学百年記念館(神大会館)六甲ホール
    • Related Report
      2011 Research-status Report
  • [Presentation] 脱イオン化ウシ血清アルブミンのマウス体細胞核移植胚への注入はその胚盤胞への発生を促進する

    • Author(s)
      伊佐治優希、西川智章、今井裕、山田雅保
    • Organizer
      日本畜産学会第115回大会
    • Place of Presentation
      名古屋大学東山キャンパス
    • Related Report
      2011 Research-status Report
  • [Book] Midkine: From Embryogenesis to Pathogenesis and Therapy.2012

    • Author(s)
      Yamada M, Isaji Y, Ikeda S
    • Total Pages
      336
    • Publisher
      Springer
    • Related Report
      2012 Research-status Report
  • [Book] 卵子学2011

    • Author(s)
      山田雅保
    • Total Pages
      1195
    • Publisher
      京都大学学術出版会
    • Related Report
      2011 Research-status Report
  • [Remarks]

    • URL

      http://www.reprod.kais.kyoto-u.ac.jp/

    • Related Report
      2013 Final Research Report

URL: 

Published: 2011-08-05   Modified: 2019-07-29  

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