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核の初期化促進方法の確立およびクローン胚から作成したntES細胞の正常性の検討

Research Project

Project/Area Number 15080211
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionThe Institute of Physical and Chemical Research

Principal Investigator

若山 照彦  The Institute of Physical and Chemical Research, ゲノム・リプログラミング研究チーム, チームリーダー (40360672)

Project Period (FY) 2003 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥108,300,000 (Direct Cost: ¥108,300,000)
Fiscal Year 2007: ¥18,000,000 (Direct Cost: ¥18,000,000)
Fiscal Year 2006: ¥18,300,000 (Direct Cost: ¥18,300,000)
Fiscal Year 2005: ¥24,000,000 (Direct Cost: ¥24,000,000)
Fiscal Year 2004: ¥24,000,000 (Direct Cost: ¥24,000,000)
Fiscal Year 2003: ¥24,000,000 (Direct Cost: ¥24,000,000)
Keywords応用動物 / 再生医学 / 獣医学 / 畜産学 / 発生・分化 / 核移植 / 初期化 / リプログラミング / クローン / ES細胞 / NT-ES細胞 / ntES細胞 / 再生医療 / 生殖細胞 / キメラ / 紡錘体
Research Abstract

我々はクローン動物および体細胞由来ntES細胞を効率よく作出する方法の開発および初期化のメカニズムの解明を試みている。昨年我々はntES細胞と受精卵由来のES細胞には違いが見られないことを証明したが、19年度はさらに詳しい比較と倫理問題の解決を試みた。我々は核移植の影響を調べるために、単為発生ES細胞を用いた遺伝子発現解析法を確立し、これによってついに我々は特定の遺伝子におけるメチル化状態について、ntES細胞とES細胞の間に違いがあることを発見した。現在このntES細胞特異的なDNAメチル化状態の違いが、その後の細胞分化や応用にどのような影響を与えるのか解析中である(Hikichi, et. al., 2007,2008)。一方倫理問題の解決については2つのアプローチを試みた。体外受精に失敗した卵子は、加齢により質が低下することから廃棄されている。この廃棄卵子を用いれば健康な女性から卵子を提供してもらうという倫理問題は解決する。そこでマウスを用いたモデル実験を行った結果、体外受精に失敗した卵子でも核移植およびntES細胞の作成には問題ないことを明らかにした(Wakayama, et. al., 2007a)。この成果は多数の全国紙やテレビで紹介された。また、胚を殺さずに1割球(細胞)を取り出し、そこからES細胞を樹立する手法を開発し、生命の萌芽である胚を壊すという問題についても回避できることを明らかにした(Wakayama, et. al., 2007b)。他に核移植技術をより簡素化した手法についての研究も報告した(Kishigami, et. al., 2007;Wakayama, et. al., 2008)。

Report

(5 results)
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (48 results)

All 2008 2007 2006 2005 2004 Other

All Journal Article (27 results) (of which Peer Reviewed: 9 results) Presentation (12 results) Book (3 results) Patent(Industrial Property Rights) (1 results) Publications (5 results)

  • [Journal Article] Effect of volume of oocyte cytoplasm on embryo development after parthenogenetic activation, intracytoplasmic sperm injection, or somatic cell nuclear transfer.2008

    • Author(s)
      Wakayama S, et. al.
    • Journal Title

      Zygote (in press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Nuclear Transfer Alters the DNA Methylation Status of Specific Genes in Fertilized And Parthenogenetically Activated Mouse Embryonic Stem Cells2008

    • Author(s)
      Hikichi T, et. al.
    • Journal Title

      Stem Cells. (in press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Efficient Strontium-Induced Activation of Mouse Oocytes in Standard Culture Media by Chelating Calcium.2007

    • Author(s)
      Kishigami and Wakayama.
    • Journal Title

      J. Reprod. Dev 53

      Pages: 1207-1215

    • NAID

      10020172730

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of progeny via ICSI following enrichment of elongated spermatids from mouse testis by flow-cytometric cell sorting.2007

    • Author(s)
      Ohta H, et. al.
    • Journal Title

      Hum Reprod. 22

      Pages: 1612-1616

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Establishment of mouse embryonic stem cell lines from somatic cell nuclei by nuclear transfer into aged, fertilization-failure mouse oocytes.2007

    • Author(s)
      Wakayama S, et. al.
    • Journal Title

      Curr. Biol. 17

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Regulation of chromatin and chromosome morphology by histone H3 modifications in pig oocytes.2007

    • Author(s)
      Bui HT, et. al.
    • Journal Title

      Reproduction. 133

      Pages: 371-382

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Efficient establishment of mouse embryonic stem cell lines from single blastomeres and polar bodies.2007

    • Author(s)
      Wakayama S, et. al.
    • Journal Title

      Stem Cells. 25

      Pages: 986-993

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Success of mouse cloning from an outbred strain by TSA treatment after somatic nuclear transfer.2007

    • Author(s)
      Kishigami S, et. al.
    • Journal Title

      J. Reprod. Dev 53

      Pages: 165-170

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Differentiation Potential of Parthenogenetic Embryonic Stem Cells in Improved by Nuclear Transfer.2007

    • Author(s)
      Hikichi T, et. al.
    • Journal Title

      Stem Cells. 25

      Pages: 46-53

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Differentiation Potential of Parthenogenetic Embryonic Stem Cells is Improved by Nuclear Transfer.2007

    • Author(s)
      Hikichi T, Wakayama S, Mizutani E, Takashima Y, Kishigami S, Van Thuan N, Ohta H, Bui HT, Nishikawa SI, Wakayama T.
    • Journal Title

      Stem Cells 25

      Pages: 46-53

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Donor Centrosome Regulation of Initial Spindle formation in Mouse Somatic Cell Nuclear Transfer : Roles of Gamma-Tubulin and Nuclear Mitotic Apparatus Protein 12006

    • Author(s)
      Van Thuan, N., Wakayama, S., Kishigami, S., Wakayama, T.
    • Journal Title

      Biol Reprod 74

      Pages: 777-787

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Injection of Somatic Cell Cytoplasm into Oocytes Before ICSI Impairs Full-Term Development and Increases Placental Weight in Mice2006

    • Author(s)
      Van Thuan, N., S.Wakayama, S.Kishigami, H.Ohta, T.Hikichi, E.Mizutani, H.T.Bui, T.Wakayama
    • Journal Title

      Biol Reprod 74

      Pages: 865-873

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Normal specification of the extraembryonic lineage after somatic nuclear transfer.2006

    • Author(s)
      Kishigami S, Hikichi T, Van Thuan N, Ohta H, Wakayama S, Bui HT, Mizutani E, Wakayama T
    • Journal Title

      FEBS Lett 580

      Pages: 1801-1806

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Equivalency of Nuclear Transfer-Derived Embryonic Stem Cells to those Derived from Fertilized Mouse Blastocyst.2006

    • Author(s)
      Wakayama S et al., (他17名)
    • Journal Title

      Stem Cells 24

      Pages: 2023-2033

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Developmental ability of cloned embryos from neural stem cells2006

    • Author(s)
      Mizutani E, Ohta H, Kishigami S, Thuan NV, Hikichi T, Wakayama S, Kosaka M, Sato E, Wakayama T
    • Journal Title

      Reproduction 132

      Pages: 849-857

    • NAID

      130007023740

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer.2006

    • Author(s)
      Kishigami S, Mizutani E, Ohta H, Hikichi T, Thuan NV, Wakayama S, Bui HT, Wakayama T.
    • Journal Title

      Biochem Biophys Res Commun 340

      Pages: 183-189

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Establishment of Male and Female Nuclear Transfer Embryonic Stem Cell Lines from Different Mouse Strains and Tissues2005

    • Author(s)
      Wakayama S, Ohta H, Kishigami S, Van Thuan N, Hikichi T, Mizutani E, Miyake M, Wakayama T
    • Journal Title

      Biol.Reprod 72

      Pages: 932-936

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Propagation of an Infertile Hermaphrodite Mouse Lacking Germ Cells, Using Nuclear Transfer and Embryonic Stem Cell Technology.2005

    • Author(s)
      Wakayama S, Kishigami S, Van Thuan N, Ohta H, Hikichi T, Muzutani E, Yanagimachi R, Wakayama T
    • Journal Title

      Proc.Natl.Acad.Sci.USA 102

      Pages: 29-33

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Generation of Normal Progeny by Intracytoplasmic Sperm Injection Following Grafting of Testicular Tissue from Cloned Mice that Died Postnatally.2005

    • Author(s)
      Ohta H, Wakayama T.
    • Journal Title

      Biol Reprod 73

      Pages: 390-395

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Generation of progeny from embryonic stem cells by microinsemination of male germ cells from chimeric mice.2005

    • Author(s)
      Mizutani E, Ohta H, Kishigami S, Van Thuan N, Hikichi T, Wakayama S, Sato E, Wakayama T
    • Journal Title

      Genesis 43

      Pages: 34-42

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Mice Cloned by Nuclear Transfer from Somatic and ntES Cells Derived from the Same Individuals2005

    • Author(s)
      Wakayama, S., E.Mizutani, S.Kishigami, N.V.Thuan, H.Ohta, T.Hikichi, H.T.Bui, M.Miyake, T.Wakayama
    • Journal Title

      J Reprod Dev 51

      Pages: 765-772

    • NAID

      10027326550

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Establishment of Male and Female Nuclear Transfer Embryonic Stem Cell Lines from Different Mouse Strains and Tissues.2005

    • Author(s)
      Wakayama et al.
    • Journal Title

      Biology of Reprodction. (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Propagation of an Infertile Hermaphrodite Mouse Lacking Germ Cells, Using Nuclear Transfer and Embryonic Stem Cell Technology.2005

    • Author(s)
      Wakayama et al.
    • Journal Title

      Proc.Natl.Acad.Sci. USA (Cover article of issue) 102

      Pages: 29-33

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Round spermatids stained with Mito Tracker can be used to produce offspring more efficiently.2005

    • Author(s)
      Hikichi et al.
    • Journal Title

      Zygote (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Similar time restriction for ICSI and ROSI into activated oocytes for efficient offspring production2004

    • Author(s)
      Kishigami et al.
    • Journal Title

      Biology of Reprodction. 70

      Pages: 1863-1869

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Skewed X-inactivation in cloned mice2004

    • Author(s)
      Senda et al.
    • Journal Title

      Biochem Biophys Res Commun 321

      Pages: 38-44

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Production of offspring from one-day-old oocytes stored at room temperature.2004

    • Author(s)
      Wakayama et al.
    • Journal Title

      Journal of Reprodction and Development. 50

      Pages: 627-637

    • NAID

      10027323645

    • Related Report
      2004 Annual Research Report
  • [Presentation] Improvement of efficiency of mouse cloning and ES cell establishment2008

    • Author(s)
      T Wakayama
    • Organizer
      The Biology of Embryonic and Adult Stem Cells
    • Place of Presentation
      Center for Cellular And Molecular Biology(ハイデルアバード市)
    • Related Report
      2007 Annual Research Report
  • [Presentation] クローン技術の問題点と応用について2007

    • Author(s)
      若山 照彦
    • Organizer
      第5回クロマチン・フロンティアーズ・ジャパン
    • Place of Presentation
      大手町サンケイプラザ
    • Year and Date
      2007-11-19
    • Related Report
      2007 Annual Research Report
  • [Presentation] From Somatic Cell to Stem Cell Through Nuclear Transfer2007

    • Author(s)
      T Wakayama
    • Organizer
      第3回日韓共同繁殖生物学シンポジウム
    • Place of Presentation
      東京大学農学部 弥生講堂
    • Year and Date
      2007-10-18
    • Related Report
      2007 Annual Research Report
  • [Presentation] クローン技術の問題点と可能性2007

    • Author(s)
      若山 照彦
    • Organizer
      再生医療の実現化プロジェクト 第4回成果発表会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-09-09
    • Related Report
      2007 Annual Research Report
  • [Presentation] Eggs that fail to become fertilized can be used as NT recipient without ethical problem2007

    • Author(s)
      T Wakayama
    • Organizer
      2007 International Symposium of Research Center For Transgenic Cloned Pigs for the title "Recent Progresses in Animal Biotechnology"
    • Place of Presentation
      Chungnam National University(ソウル市)
    • Year and Date
      2007-09-06
    • Related Report
      2007 Annual Research Report
  • [Presentation] Eggs that fail to become fertilized can be used as N recipient without ethical problem2007

    • Author(s)
      T Wakayama
    • Organizer
      Present and Future of Transgenic Cloned Pig Researth
    • Place of Presentation
      Chungnam National University(チョングナム市)
    • Year and Date
      2007-09-06
    • Related Report
      2007 Annual Research Report
  • [Presentation] マウスの遺伝子保全方法の開発2007

    • Author(s)
      若山 照彦
    • Organizer
      第4回理化学研究所 実験動物懇談会
    • Place of Presentation
      理化学研究所 筑波研究所
    • Related Report
      2007 Annual Research Report
  • [Presentation] 受精失敗加齢卵子を用いたICSI,NTおよびntES細胞について2007

    • Author(s)
      若山 清香・若山 照彦
    • Organizer
      第48回日本哺乳動物卵子学会
    • Place of Presentation
      アピオ甲府
    • Related Report
      2007 Annual Research Report
  • [Presentation] 倫理問題のない核移植技術の開発2007

    • Author(s)
      若山 照彦
    • Organizer
      第25回日本ヒト細胞学会
    • Place of Presentation
      都市センターホテル
    • Related Report
      2007 Annual Research Report
  • [Presentation] Improvement of the efficiency of somatic cell nuclear transfer2007

    • Author(s)
      T Wakayama
    • Organizer
      "Therapeutic Cloning": Where Do We Go From Here?
    • Place of Presentation
      National Academies(アーバイン市)
    • Related Report
      2007 Annual Research Report
  • [Presentation] 再クローニングの再チャレンジ2007

    • Author(s)
      若山 照彦
    • Organizer
      「生殖細胞の発生プロセス・再プログラム化とエピジェネティクス」
    • Place of Presentation
      コクヨホール
    • Related Report
      2007 Annual Research Report
  • [Presentation] Nuclear reprogramming to produce cloned mice and ES cell from somatic cells2007

    • Author(s)
      T Wakayama
    • Organizer
      Asian Reproductive Biotechnology Society
    • Place of Presentation
      National Universiyt of Singapore(シンガポール市)
    • Related Report
      2007 Annual Research Report
  • [Book] A laboratory handbook, Third edition. Vol. 1 (ed Celis JE)2007

    • Author(s)
      Wakayama T.
    • Publisher
      Academic press, San Diego, California, USA
    • Related Report
      2007 Annual Research Report
  • [Book] In Cell Biology : A laboratory handbook, Third edition. Vol. 12006

    • Author(s)
      Wakayama T(編集Celis JE)
    • Total Pages
      586
    • Publisher
      Academic press, San Diego, California, USA
    • Related Report
      2006 Annual Research Report
  • [Book] Genetically Engineered Mice (Mouse genetic resources without germ cells ; somatic cell nuclear transfer and ES technology) (eds Sundberg.J.P. & Ichiki T.)2005

    • Author(s)
      Wakayama T
    • Publisher
      CRC Press, Boca Raton, USA
    • Related Report
      2005 Annual Research Report
  • [Patent(Industrial Property Rights)] 非ヒトES動物の新規作成方法2007

    • Inventor(s)
      大田浩、坂出裕子、山縣一夫、若山照彦
    • Industrial Property Rights Holder
      独立行政法人理化学研究所
    • Industrial Property Number
      2007-171788
    • Filing Date
      2007-06-29
    • Related Report
      2007 Annual Research Report
  • [Publications] Wakayama S: "Effect of timing of the removal of oocyte chromosomes before or after injection of somatic nucleus on development of NT embryos"Cloning and stem cells. 5. 181-189 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tamashiro KL: "Phenotype of Cloned Mice : Development, Behavior, and Physiology."Exp Biol Med. (In press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Singh U: "Abnormal gene expression in mouse placentopathies caused by interspecies hybridization, cloning, and Esx1 mutation"Developmental Dynamics. (In press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yanagimachi R: "Production of fertile offspring from genetically infertile male mice"Proc.Natl.Acad.Sci.USA. (In press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Lanza R: "Regeneration of the Infarcted Heart With Stem Cells Derived by Nuclear Transplantation"Circulation Research. (In press). (2004)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2018-03-28  

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