Regulatory mechanism of mouse GSC/niche system and in vitro spermatogenesis

2012 Fiscal Year Final Research Report

• Project Area: Regulatory Mechanism of Gamete Stem Cells
• Project/Area Number: 20116005
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Yokohama City University
• Principal Investigator: OGAWA Takehiko 横浜市立大学, 医学研究科, 准教授 (50254222)
• Co-Investigator(Kenkyū-buntansha): OHBO Kazuyuki 横浜市立大学, 医学研究科, 教授 (70250751)
• Project Period (FY): 2008 – 2012
• Keywords: 精子形成 / 精子幹細胞 / 器官培養

Research Abstract

We succeeded to produce mouse sperm from spermatogonial stem cells (SSCs) in vitro. We adopted an organ culture method, and found that serum replacement was effective to induce complete spermatogenesis in our system. When cultured SSCs were transplanted into the seminiferous tubules of extracted host testes, they also differentiated into sperm in the cultured testis tissues. In addition, we demonstrated that an infertile model mouse, Sl/Sl^d mutant, can be treated with the explantation of their testis tissues.

Research Products

• [Journal Article] In vitro spermatogenesis using an organ culture technique (2013)
  • Author(s): T. Yokonishi, T. Sato, K. Katagiri, T. Ogawa
  • Journal Title: Methods Mol Biol Volume: 927 Pages: 479-488
• [Journal Article] Testis tissue explantation cures spermatogenic failure in c-Kit ligand mutant mice. (2012)
  • Author(s): T. Sato, T. Yokonishi, M. Komeya, K. Katagiri, Y. Kubota, S. Matoba, N. Ogonuki, A. Ogura, S. Yoshida, T. Ogawa
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Pages: 16934-16938
  • DOI: DOI:10.1073/pnas.1211845109
  • Peer Reviewed
• [Journal Article] Splice variant HE4-V3 expression is associated with favorable prognosis in pulmonary adenocarcinoma (2012)
  • Author(s): K. Tokuishi, SI. Yamashita, K. Ohbo and K. Kawahara
  • Journal Title: Tumour Biol Volume: 33 Pages: 103-109
  • DOI: doi:10.1007/s13277-011-0252-8
  • Peer Reviewed
• [Journal Article] Serum level of HE4 is closely associated with pulmonary adenocarcinoma progression (2012)
  • Author(s): SI. Yamashita, K. Tokuishi, T. Moroga, S. Yamamoto, K. Ohbo, S. Miyahara, Y. Yoshida, J. Yanagisawa, D. Hamatake, M. Hiratsuka, Y. Yoshinaga, T. Shiraishi, A. Iwasaki and K. Kawahara
  • Journal Title: Tumour Biol Volume: 33 Pages: 2365-2370
  • DOI: doi:10.1007/s13277-012-0499-8
  • Peer Reviewed
• [Journal Article] In vitro production of fertile sperm from murine spermatogonial stem cell lines (2011)
  • Author(s): T. Sato, K. Katagiri, T. Yokonishi, Y. Kubota, K. Inoue, N. Ogonuki, S. Matoba, A. Ogura, T. Ogawa
  • Journal Title: Nat Commun Volume: 2 Pages: 472
  • DOI: doi:10.1038/ncomms1478
  • Peer Reviewed
• [Journal Article] In vitro production of functional sperm in cultured neonatal mouse testes (2011)
  • Author(s): T. Sato, K. Katagiri, A. Gohbara, K. Inoue, N. Ogonuki, A. Ogura, Y. Kubota, T. Ogawa
  • Journal Title: Nature Volume: 471 Pages: 504-507
  • DOI: doi:10.1038/nature09850
  • Peer Reviewed
• [Journal Article] HP1g links histone methylation marks to meiotic synapsis in mice (2011)
  • Author(s): Y. Takada Y, C. Naruse, Y. Costa, T. Shirakawa, M. Tachibana, J. Sharif, F. Kezuka-Shiotani, D. Kakiuchi, H. Masumoto, Y. Shinkai, K. Ohbo, A. H.F.M. Peters, J. A.M. Turner, M. Asano and K. Koseki
  • Journal Title: Development Volume: 138 Pages: 4207-4217
  • DOI: doi:10.1242/dev.064444
  • Peer Reviewed
• [Journal Article] Proliferation of Mouse Spermatogonial Stem Cells in Microdrop Culture (2010)
  • Author(s): Y. Araki, T. Sato, K. Katagiri, Y. Kubota, Y. Araki, T. Ogawa
  • Journal Title: Biol Reprod Volume: 83 Pages: 951-957
  • DOI: doi:10.1095/ biolreprod.109.082800
  • Peer Reviewed
• [Journal Article] In Vitro Murine Spermatogenesis in an Organ Culture System (2010)
  • Author(s): A. Gohbara, K. Katagiri, T. Sato, Y. Kubota, H. Kagechika, Y. Araki, Y. Araki, T. Ogawa
  • Journal Title: Biol Reprod Volume: 83 Pages: 261-267
  • DOI: doi:10.1095/biolreprod.110.083899
  • Peer Reviewed
• [Journal Article] Ectopic porcine spermatogenesis in murine subcutis: tissue grafting versus cell-injection methods (2009)
  • Author(s): T. Watanabe, H. Hayashi, K. Kita, Y. Kubota, T. Ogawa
  • Journal Title: Asian J Androl Volume: 11 Pages: 317-323
  • Peer Reviewed
• [Journal Article] The histone3 lysine 4 methyltransferase, Mll2, is only required briefly in development and spermatogenesis (2009)
  • Author(s): Glaser S., Lubitz S., Loveland KL., Ohbo K., Robb L., Schwenk F., Seibler J., Roellig D, Kranz A., Anastassiadis K. and Stewart AF
  • Journal Title: Epigenetics & Chromatin Volume: 2 Pages: 5
  • DOI: doi:10.1186/1756-8935-2-5
  • Peer Reviewed
• [Presentation] In vitro spermatogenesis using an organ culture method (2013)
  • Author(s): T. Ogawa
  • Organizer: Australia Melbourne 10th International Congress of Andrology 10th International Congress of Andrology
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20130223-26
• [Presentation] 精子形成研究の変遷と展望 (2013)
  • Author(s): 小川毅彦
  • Organizer: 第101回日本泌尿器科学会総会
  • Place of Presentation: さっぽろ芸術文化の館 (北海道)
  • Year and Date: 2013-04-28
• [Presentation] 無精子症治療の新たな展開 - ヒト精細胞培養法の開発 (2013)
  • Author(s): 小川毅彦
  • Organizer: 第101回日本泌尿器科学会総会 シンポジウム5 新しい治療戦略シリーズ 男性不妊症診療の新たな戦略
  • Place of Presentation: ロイトン札幌 (北海道)
  • Year and Date: 2013-04-28
• [Presentation] In vitro reconstruction of functional mouse seminiferous tubules supporting germ cell differentiation 培養下での精巣組織再構成の試み (2013)
  • Author(s): T. Ogawa
  • Organizer: 第90回 日本生理学会大会
  • Place of Presentation: タワーホール船堀(東京)
  • Year and Date: 2013-03-29
• [Presentation] In vitro reconstruction of functional testicular tissues supporting spermatogenesis (2013)
  • Author(s): T. Ogawa
  • Organizer: 日本再生医療学会 Johnson & Johnson Innovation Award
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2013-03-22
• [Presentation] In vitro spermatogenesis using an organ culture method (2012)
  • Author(s): T. Ogawa
  • Organizer: Meeting GDRI "Mammalian Meiosis Network"
  • Place of Presentation: La Maison du Seminaire, Nice, France
  • Year and Date: 20121011-12
• [Presentation] 精巣組織培養法は無精子症を治療できるかシンポジウム2「無精子症の取り扱いに関する問題と展望」 (2012)
  • Author(s): 小川毅彦
  • Organizer: 日本生殖医学会学術講演会
  • Place of Presentation: 長崎ブリックホール(長崎県)
  • Year and Date: 2012-11-08
• [Presentation] Induction of spermatogenesis in testis of c-Kit ligand mutant spermatogenic failure mice using Organ Culture method (2012)
  • Author(s): T.Ogawa
  • Organizer: 63rd Congress of Korean Society for Reproductive Medicine
  • Place of Presentation: Seoul Women's University(Korea)
  • Year and Date: 2012-11-01
• [Presentation] 精子幹細胞からの培養下精子形成 (2012)
  • Author(s): 小川毅彦
  • Organizer: 日本受精着床学会シンポジウム「基礎(1)精子幹細胞のバイオロジーとその応用」
  • Place of Presentation: 大阪国際会議場(大阪府)
  • Year and Date: 2012-08-30
• [Presentation] Changing epigenetic modification and chromocenter formation associating with spermatogonia differentiation (2012)
  • Author(s): K. Ohbo, T. Shirakawa and H. Koseki
  • Organizer: 10^<th> .EMBL Conference, Transcription and Chromatin
  • Place of Presentation: Heidelbeerg, Germany
  • Year and Date: 2012-08-26
• [Presentation] In vitro spermatogenesis using an organ culture method (2012)
  • Author(s): T. Ogawa
  • Organizer: The 58/60th NIBB symposium
  • Place of Presentation: Okazaki, (Aichi prefecture) Japan
  • Year and Date: 2012-07-21
• [Presentation] The 58th/60th NIBB Conference, Germline Specification, Sex, and Stem Cells (2012)
  • Author(s): K. Ohbo
  • Organizer: Analysis of epigenetic changes associated with spermatogonia stem cell differentiation
  • Place of Presentation: Okazaki, (Aichi prefecture) Japan
  • Year and Date: 2012-07-19
• [Presentation] State-of-the-Art Lecture 1 "Testicular tissue culturing inducing complete spermatogenesis" (2012)
  • Author(s): T. Ogawa
  • Organizer: The 7th AUA/JUA international affiliate society meeting
  • Place of Presentation: Georgia World Congress Center, Atlanta
  • Year and Date: 2012-05-20
• [Presentation] In vitro精子形成法の開発と不妊症診療への応用の可能性 (2012)
  • Author(s): 小川毅彦
  • Organizer: 第100回 日本泌尿器科学会総会SY2-5「男性不妊症に対する新たな挑戦」
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2012-04-22
• [Presentation] 器官培養法を用いたin vitro精子形成法の開発 (2012)
  • Author(s): 小川毅彦
  • Organizer: 第20回平成不妊研究会
  • Place of Presentation: 六本木・アカデミーヒルズ(東京都)
  • Year and Date: 2012-03-15
• [Presentation] Analysis of epigenetic feature of spermatogonial stem cells in mice (2012)
  • Author(s): K. Ohbo, T. Shirakawa, R. Yaman- Deveci, Y. Kamizato, K. Nakajima, H. Sone, Y. Sato, S. Jafar, Y. Takada-Horisawa, S. Yoshida, K. Ura, M. Muto and H. Koseki
  • Organizer: Keystone Symposia, Epigenomics, Chromatin Dynamics
  • Place of Presentation: Colorado, USA
  • Year and Date: 2012-01-19
• [Presentation] 器官培養法を用いたin vitro精子形成法の開発 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第26回動物生殖工学研究会
  • Place of Presentation: 北里大学・北里本館2F 大会議室(東京都)
  • Year and Date: 2011-12-04
• [Presentation] In vitro精子形成法の開発と臨床応用の可能性 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第3回生殖補助医療胚培養士セミナー
  • Place of Presentation: JA共済ビル(東京都)
  • Year and Date: 2011-11-13
• [Presentation] 器官培養法を用いた精子形成法の開発と臨床応用の可能性 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第2回生殖医療研究会
  • Place of Presentation: 国際医療福祉大学 東京青山キャンパス(東京都)
  • Year and Date: 2011-11-06
• [Presentation] 器官培養法を用いたin vitro精子形成法の開発と発展 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第89回発生工学・疾患モデル研究会
  • Place of Presentation: 順天堂大学10号館8F803(東京都)
  • Year and Date: 2011-10-26
• [Presentation] 器官培養法によるin vitro精子形成と臨床応用の可能性 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第76回日本泌尿器科学会東部総会 シンポジウム
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2011-10-22
• [Presentation] In vitro spermatogenesis (2011)
  • Author(s): T. Ogawa
  • Organizer: 57th Annual meeting of Canadian Fertility and Andrology Society
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2011-09-21
• [Presentation] マウス精巣の器官培養によるin vitro精子形成 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第104回日本繁殖学会シンポジウム「精原幹細胞:基礎から応用まで」
  • Place of Presentation: いわて県民情報交流センター・アイーナ(岩手県)
  • Year and Date: 2011-09-17
• [Presentation] In vitro production of functional sperm in cultured neonatal mouse testes (2011)
  • Author(s): T. Ogawa
  • Organizer: 16th World Congress on In vitro Fertilization & 5th World Congress on In vitro Maturation
  • Place of Presentation: Keio Plaza (Tokyo), Japan
  • Year and Date: 2011-09-13
• [Presentation] Identification of an epigenetic checkpoint that controls the differentiation from spermatogonial stem cells to progenitor cells (2011)
  • Author(s): T. Shirakawa, R. Yaman-Deveci, Y. Kamizato, K. Nakajima, H. Sone, Y. Sato, S. Jafar, Y. Takada-Horisawa, S. Yoshida, K. Ura, M. Muto, H. Koseki, T. Suda and K. Ohbo
  • Organizer: Stem Cells in Development and Disease
  • Place of Presentation: Berlin, Germany
  • Year and Date: 2011-09-12
• [Presentation] An epigenetic checkpoint controls the transition from a stem cell pool to a progenitor cell stage in testes (2011)
  • Author(s): K. Ohbo
  • Organizer: EMBO Conference Series on Chromatin and Epigenetics
  • Place of Presentation: EMBL, Heidelberg, Germany
  • Year and Date: 2011-06-04
• [Presentation] 器官培養法を用いた新生仔マウス精巣でのin vitro精子産生 (2011)
  • Author(s): 小川毅彦、佐藤卓也、横西哲広、郷原絢子、河路かおる、窪田吉信
  • Organizer: 第99回 日本泌尿器科学会総会
  • Place of Presentation: 名古屋国際会議場(愛知県)
  • Year and Date: 2011-04-22
• [Presentation] in vitro精子形成の進展 (2011)
  • Author(s): 小川毅彦
  • Organizer: 第6回 日本生殖再生医学会
  • Place of Presentation: シェーンバッハ・サボー(東京都)
  • Year and Date: 2011-03-13
• [Presentation] 精子幹細胞の培養とin vitro 精子形成の進展 (2010)
  • Author(s): 小川毅彦
  • Organizer: 第55回日本生殖医学会総会/シンポジウム3「精子形成研究のあらたなるフロンティア: 幹細胞から精子の品質まで」
  • Place of Presentation: 徳島市あわぎんホール(徳島県)
  • Year and Date: 2010-11-12
• [Presentation] In vitro production of functional sperm in cultured neonatal mouse testes (2010)
  • Author(s): T. Ogawa, T. Sato, K. Katagiri, Y. Kubota
  • Organizer: Cold Spring Harbor Symposium on Germ Cells
  • Place of Presentation: NewYork, USA
  • Year and Date: 2010-10-08
• [Presentation] Epigenetic difference between stem cells and progenitor cells in testes (2010)
  • Author(s): K. Ohbo
  • Organizer: Advances in Epigenetics Epigenome NoE and Chromatin Plasticity
  • Place of Presentation: San Feliu deGuixols, France
  • Year and Date: 2010-05-13
• [Presentation] エピジェネティクスの視点から見た、精巣幹細胞維持と前駆細胞への分化メカニズム (2010)
  • Author(s): 大保和之,白川峰征、中島久仁子
  • Organizer: 第115回日本解剖学会総会
  • Place of Presentation: 盛岡・岩手県民会館(岩手県)
  • Year and Date: 2010-03-30
• [Presentation] 培養精子幹細胞(GS細胞)への効率的な遺伝子導入 (2010)
  • Author(s): 佐藤卓也、片桐久美子、窪田吉信、小川毅彦
  • Organizer: 第5回日本生殖再生医学会学術集会
  • Place of Presentation: シェーンバッハ・サボー(東京都)
  • Year and Date: 2010-02-21
• [Presentation] 精原幹細胞と精子形成 (2010)
  • Author(s): 小川毅彦、佐藤卓也 、片桐久美子、窪田吉信、荒木泰行、荒木康久
  • Organizer: 第13回 RMB研究会シンポジウム
  • Place of Presentation: 持田製薬株式会社「ルークホール」(東京都)
  • Year and Date: 2010-01-23
• [Presentation] マウス精子幹細胞のマイクロドロップ培養下での増殖動態 (2009)
  • Author(s): 小川毅彦、荒木泰行、荒木康久
  • Organizer: 第41回精子研究会
  • Place of Presentation: 東京大学 駒場キャンパス 16号館 119-129室(東京都)
  • Year and Date: 2009-12-19
• [Presentation] 精子幹細胞の培養と精子形成 (2009)
  • Author(s): 小川毅彦
  • Organizer: 第14回日本生殖内分泌学会学術集会 シンポジウム-生殖臓器由来幹細胞研究の最前線
  • Place of Presentation: シェーンバッハ・サボー(東京都)
  • Year and Date: 2009-11-28
• [Presentation] Difference of epigenetic modifications between self-renewing tissue-specific stem cells and progenitor cells in mouse testes (2009)
  • Author(s): Shikarawa T, Ura K, Yoshida S, Ohbo K
  • Organizer: EMBO Conference Series on Nuclear Structure and Dynamics
  • Place of Presentation: Isle sur la Sorgue, France
  • Year and Date: 2009-10-03
• [Presentation] in vitro にもニッチはあるか? (2009)
  • Author(s): 小川毅彦
  • Organizer: 第80回 日本動物学会・シンポジウム「配偶子幹細胞制御に関する研究の新展開」
  • Place of Presentation: 静岡グランシップ(静岡県)
  • Year and Date: 2009-09-17
• [Presentation] Dynamic epigenetic changes in differentiation step from spermatogonial stem cells to progenitor cells (2009)
  • Author(s): Shirakawa T, Ohbo K
  • Organizer: Gordon Research Conferences Mammalian Gametogenesis & Embyogenesis
  • Place of Presentation: Waterville Valley, NH, USA
  • Year and Date: 2009-08-03
• [Presentation] In vitro murine spermatogenesis in an organ culture system (2009)
  • Author(s): T. Ogawa, K. Katagiri, Y. Araki, Y. Araki, Y. Kubota
  • Organizer: European Society of Human Reproduction and Embryology (ESHRE)
  • Place of Presentation: Amsterdam, Nederland
  • Year and Date: 2009-07-01
• [Presentation] 精巣幹細胞特異的発現分子により可視化された幹細胞のゲノム修飾動態 (2009)
  • Author(s): 大保 和之
  • Organizer: シンポジウム「生殖系細胞の機能発現とイメージング」第114回日本解剖学会総会・全国学術総会
  • Place of Presentation: 岡山理科大学(岡山県)
  • Year and Date: 2009-03-28
• [Presentation] 精巣幹細胞から前駆細胞への分化に伴う遺伝子発現制御機構の変化 (2009)
  • Author(s): 大保和之
  • Organizer: SSRE生殖工学研究会シンポジウム「生殖幹細胞の分化制御機構」
  • Place of Presentation: 明治大学駿河台校舎(東京都)
  • Year and Date: 2009-03-07
• [Remarks]
  • URL: http://www-user.yokohama-cu.ac.jp/~urology/