家畜精巣由来の生殖幹細胞培養と生殖細胞への分化制御に関する研究

• 研究課題/領域番号: 22380150
• 研究種目: 基盤研究(B)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 応用動物科学
• 研究機関: 京都大学
• 研究代表者: 今井 裕 京都大学, 大学院・農学研究科, 教授 (10303869)
• 研究期間 (年度): 2010 – 2012
• 研究課題ステータス: 完了 (2012年度)
• 配分額: 19,760千円 (直接経費: 15,200千円、間接経費: 4,560千円); 2012年度: 4,550千円 (直接経費: 3,500千円、間接経費: 1,050千円); 2011年度: 6,500千円 (直接経費: 5,000千円、間接経費: 1,500千円); 2010年度: 8,710千円 (直接経費: 6,700千円、間接経費: 2,010千円)
• キーワード: 発生工学 / 生殖幹細胞

研究概要

ウシ精巣に由来する前精原幹細胞の性質と体外での長期培養について検討した。本研究によって、ウシの前精原幹細胞は、無血清培地下でポリLリシンを細胞接着基質とする培養条件下で2 カ月以上にわたってコロニーの増殖と生存性を維持することが可能になった。また、これらのコロニーは、多能性分化能と精原幹細胞としての性質を併せ持っていた。長期培養用培地に成長因子を添加して培養すると、減数分裂直前の精母細胞に分化させることができた。

研究成果

• [雑誌論文] Effects of extracellular matrices and lectin Dolichos biflorus agglutinin on cell adhesion and self-renewal of bovine gonocytes cultured in vitro (2013)
  • 著者名/発表者名: Kim SM, Fujihara M, Sahare M, Minami N, Yamada M and Imai H
  • 雑誌名: Reproduction Fertility and Development 巻: In press 号: 2 ページ: 268-268
  • DOI: 10.1071/rd12214
  • NAID: 120005367805
  • 査読あり
• [雑誌論文] Protein Kinase A Determines Timing of Early Differentiation through Epigenetic Regulation with G9a (2012)
  • 著者名/発表者名: Yamamizu K, Fujihara M, Tachibana M, Katayama S, Takahashi A, Hara E, Imai H, Shinkai Y, and Yamashita JK
  • 雑誌名: Cell Stem Cell 巻: 10 号: 6 ページ: 759-770
  • DOI: 10.1016/j.stem.2012.02.022
  • 査読あり
• [雑誌論文] 哺乳動物におけるiPS細胞 (2012)
  • 著者名/発表者名: 浅野良太
  • 雑誌名: 日本胚移植研究会誌 巻: 34 ページ: 9-18
  • 査読あり
• [雑誌論文] Simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system (2011)
  • 著者名/発表者名: Tsukiyama T., Asano R., Kawaguchi T., Kim N., Yamada M., Minami N., Ohinata Y. and Imai H
  • 雑誌名: Genes to Cells 巻: 16 号: 7 ページ: 815-825
  • DOI: 10.1111/j.1365-2443.2011.01528.x
  • NAID: 120005593680
  • 査読あり
• [雑誌論文] Characterization and in vitro culture of male germ cells from developing bovine testes (2011)
  • 著者名/発表者名: Fujihara M, Kim SM, Minami N, Yamada M and Imai H
  • 雑誌名: Journal of Reproduction and Development 巻: 57 ページ: 355-364
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/21289464
  • 査読あり
• [雑誌論文] Identification of reprogramming factors associated with a pluripotent potential of porcine and bovine spermatic stem cells in culture (2011)
  • 著者名/発表者名: Kim SM, Fujihara M, Goel S, Sahare M, Minami N, Yamada M and Imai H
  • 雑誌名: The Proceedings for 16th World Congress on In Vitro Fertilization ページ: 41-48
  • URL: http://isivf2011.org/jp/
  • 査読あり
• [雑誌論文] UCHL-1 protein expression specifically marks spermatogonia in wild bovid testis (2011)
  • 著者名/発表者名: Goel S, Mahla RS, Suman SK, Reddy N and Imai H
  • 雑誌名: European Journal of Wildllife Research 巻: 57 号: 3 ページ: 663-667
  • DOI: 10.1007/s10344-010-0454-1
• [雑誌論文] Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos (2011)
  • 著者名/発表者名: Miyamoto, K., Nagai, K., Kitamura, N., Nishikawa, T., Ikegami, H., Binh, N., Tsukamoto, S., Matsumoto, M., Tsukiyama, T., Minami, N., Yamada, M., Ariga, H., Miyake, M., Kawarasaki, T., Matsumoto, K., Imai, H.
  • 雑誌名: Proc. Natl. Acad. Sci. USA 巻: 108 号: 17 ページ: 7040-7045
  • DOI: 10.1073/pnas.1013634108
  • 査読あり
• [雑誌論文] Pluripotent stem cells from testis (2011)
  • 著者名/発表者名: Goel S., Imai H.
  • 雑誌名: Embryonic Stem Cells-Differentiation and Pluripotent Alternatives(ed.Michael S.Kallos, InTech Publisher, Rijeka) ページ: 473-492
  • 査読あり
• [雑誌論文] Identification of reprogramming factors associated with a pluripotent potential of porcine and bovine spermatic stem cells in culture (2011)
  • 著者名/発表者名: Kim SM, Fujihara M, Goel S, Sahare M, Minami N, Yamada M, Imai H
  • 雑誌名: The Proceedings for 16th World Congress on In Vitro Fertilization ページ: 41-48
  • 査読あり
• [雑誌論文] Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos. (2011)
  • 著者名/発表者名: 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.
  • 雑誌名: Proceedings of the National Academy of Science USA 巻: (印刷中)
  • 査読あり
• [雑誌論文] A simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system. (2011)
  • 著者名/発表者名: Tsukiyama T, Asano R, Kawaguchi T, Kim N, Yamada M, Minami N, Ohinata Y, Imai H.
  • 雑誌名: Genes to Cells 巻: (印刷中)
  • NAID: 120005593680
  • 査読あり
• [雑誌論文] Characterization and in vitro culture of male germ cells from developing bovine testis (2011)
  • 著者名/発表者名: Fujihara M., Kim SM., Minami N, Yamada M., Imai H.
  • 雑誌名: Journal of Reproduction and Development 巻: (印刷中)
  • NAID: 10029050501
  • 査読あり
• [雑誌論文] UCHL-1 protein expression specifically marks spermatogonia in wild bovid testis. (2011)
  • 著者名/発表者名: Goel S, Mahla RS, Suman SK, Reddy N, Imai H.
  • 雑誌名: European Journal of Wild Animal Research 巻: (印刷中)
  • 査読あり
• [雑誌論文] Spermatogonia-specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia (2010)
  • 著者名/発表者名: Goel S, Reddy N, Mandal S, Fujihara M, Kim SM and Imai H
  • 雑誌名: Theriogenology 巻: 74 号: 7 ページ: 1221-1232
  • DOI: 10.1016/j.theriogenology.2010.05.025
  • 査読あり
• [雑誌論文] Spermatogonia specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia. (2010)
  • 著者名/発表者名: Goel S., Reddy N., Mandal S., Fujihara M., Kim SM., Imai H.
  • 雑誌名: Theriogenology 巻: 74 ページ: 1221-1232
  • 査読あり
• [学会発表] Long term self-renewal and proliferation of bovine male germ stem cells in vitro (2013)
  • 著者名/発表者名: Sahare M, Kim SM, Otomo A, Komatsu K, Minami N, Yamada M and Imai H
  • 学会等名: CiRA International Symposium 2013 ~Raising the next generation of stem cell research
  • 発表場所: Kyoto
• [学会発表] Long term self-renewal and proliferation of bovine male germ stem cells in vitro. (2013)
  • 著者名/発表者名: 今井 裕
  • 学会等名: CiRA International Symposium 2013 ~Raising the next generation of stem cell research~
  • 発表場所: Kyoto, Japan
• [学会発表] Mitogen activated protein kinase (MAPK) signaling is indispensable for maintaning self renewal and proliferation of bovine male germ stem cells in culture (2013)
  • 著者名/発表者名: 今井 裕
  • 学会等名: The 2013 Society for Study of Reproduction (SSR) Annual Meeting
  • 発表場所: Montreal, Canada
• [学会発表] Glycan epitopes involve in germ cell differentiation during early development of neonatal mice testis (2012)
  • 著者名/発表者名: Kim SM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • 学会等名: The 4th Congress of the Asia Pacific Initiative on Reproduction
  • 発表場所: Osaka
• [学会発表] The role of GalNAc glycan residues associated adhesion and self-renewal in cultured bovine gonocytes (2012)
  • 著者名/発表者名: Kim SM, Fujihara M, Sahare M, Minami N, Yamada M and Imai H
  • 学会等名: The 17th International Congress on Animal Reproduction (ICAR)
  • 発表場所: Vancouver, Canada
• [学会発表] Determination of distinct germ cell commitment by the specific affinity of the lectin DBA during early development of gonocytes in neonatal mice testis (2012)
  • 著者名/発表者名: KimSM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • 学会等名: Symposium on Functional Genomics of Early Livestock Development
  • 発表場所: Banff, Canada
• [学会発表] Glycan epitopes involve in germ cell differentiation during early development of neonatal mice testis. (2012)
  • 著者名/発表者名: 今井 裕
  • 学会等名: The 4th Congress of the Asia Pacific Initiative on Reproduction
  • 発表場所: Osaka, Japan
• [学会発表] The role of GalNAc glycan residues associated adhesion and self-renewal in cultured bovine gonocytes. (2012)
  • 著者名/発表者名: 今井 裕
  • 学会等名: The 17th International Congress on Animal Reproduction (ICAR).
  • 発表場所: Vancouver, Canada
• [学会発表] Determination of distinct germ cell commitment by the specific affinity of the lectin DBA during early development of gonocytes in neonatal mice testis. (2012)
  • 著者名/発表者名: 今井 裕
  • 学会等名: Symposium on Functional Genomics of Early Livestock Development
  • 発表場所: Banff, Canada
• [学会発表] 家畜ならびに大型実験動物における多能性幹細胞株の樹立と応用 (2010)
  • 著者名/発表者名: 築山智之・今井 裕
  • 学会等名: 日本生化学会九州支部例会シンポジウム(招待講演)
  • 発表場所: 鹿児島市
  • 年月日: 2010-05-23
• [学会発表] 家畜ならびに大型実験動物における多能性幹細胞株の樹立と応用 (2010)
  • 著者名/発表者名: 築山智之・今井裕
  • 学会等名: 日本生化学会九州支部例会シンポジウム
  • 発表場所: 鹿児島大学(鹿児島)(招待講演)
  • 年月日: 2010-05-23
• [学会発表] Comparative Changes on Stem Cell Potential of Bovine Male Germ Cells During Testicular Development from Neonatal to Adult (2010)
  • 著者名/発表者名: Fujihara M, Kim SM, Minami N, Yamada M and Imai H
  • 学会等名: The 43rd Annual Meeting of the Study of the Society for Reproduction (SSR)
  • 発表場所: Milwaukee USA
• [学会発表] Stem cells of the male germline: an aid to conservation of endangered species (2010)
  • 著者名/発表者名: Goel S, Mandal S, Reddy N, Mahara R, Suman S, Raj TA, Fujihara M, Kim SM and Imai H
  • 学会等名: The 11th International Symposium on Spermatology (ISS)
  • 発表場所: Okinawa, Japan
• [学会発表] Lectin DBA specifically recognizes glycan cell surface epitope of gonocytes in developing mouse testis (2010)
  • 著者名/発表者名: Kim SM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • 学会等名: The 11th International Symposium on Spermatology (ISS)
  • 発表場所: Okinawa, Japan
• [学会発表] Analysis of culture conditions for generation of germ-line competent induced pluripotent stem (iPS) cells in various mammalian species (2010)
  • 著者名/発表者名: Tsukiyama T, Asano R, Minami N, Yamada M and Imai H
  • 学会等名: The 8th Annual Meeting of the International Society for Stem Cell Research
  • 発表場所: San Francisco USA
• [学会発表] Comparative Changes on Stem Cell Potential of Bovine Male Germ Cells During Testicular Development from Neonatal to Adult (2010)
  • 著者名/発表者名: Fujihara M, Kim SM, Minami N, Yamada M, Imai H.
  • 学会等名: The 43rd Annual Meeting of the Study of the Society for Reproduction (SSR)
  • 発表場所: USA(Milwaukee)
• [学会発表] Stem cells of the male germline : an aid to conservation of endangered species. (2010)
  • 著者名/発表者名: Goel S, Mandal S, Reddy N, Mahara R, Suman S, Raj TA, Fujihara M, Kim SM, Imai H.
  • 学会等名: The 11th International Symposium on Spermatology(ISS)
  • 発表場所: 沖縄コンベンションセンター
• [学会発表] Lectin DBA specifically recognizes glycan cell surface epitope of gonocytes in developing mouse testis. (2010)
  • 著者名/発表者名: Kim SM, Fujihara M, Goel S, Minami N, Yamada M, Imai H.
  • 学会等名: The 11th International Symposium on Spermatology(ISS)
  • 発表場所: 沖縄コンベンションセンター
• [学会発表] Analysis of culture conditions for generation of germ-line competent induced pluripotent stem(iPS)cells in various mammalian species. (2010)
  • 著者名/発表者名: Tsukiyama T, Asano R, Minami N, Yamada M, Imai H.
  • 学会等名: The 8^<th> Annual Meeting of the International Society for Stem Cell Research
  • 発表場所: USA(San Francisco)
• [図書] Pluripotent stem cells from testis."Embryonic Stem Cells - Differentiation and Pluripotent Alternatives" (2011)
  • 著者名/発表者名: Goel S. and Imai H
  • 出版者: ed. Michael S. Kallos、InTech Publisher、Rijeka
• [備考]
  • URL: http://www.reprod.kais.kyoto-u.ac.jp/index.htm
• [備考] 京都大学大学院農学研究科応用生物科学専攻生殖生物学分野
  • URL: http://www.reprod.kais.kyoto-u.ac.jp/
• [備考]
  • URL: http://www.reprod.kais.kyoto-u.ac.jp/index.htm