Long term culture and regulation of differentiation of germ cells from the testis in domestic species

• Project/Area Number: 22380150
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: Kyoto University
• Principal Investigator: IMAI Hiroshi 京都大学, 大学院・農学研究科, 教授 (10303869)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥19,760,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥4,560,000); Fiscal Year 2012: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2011: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2010: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
• Keywords: 発生工学 / 生殖幹細胞

Research Abstract

A long-term culture of male primitive germ cells from bovine testis has been examined. Bovine primitive germ cells can proliferate and survive in culture for over 2 months in the condition with serum-free components and on poly-L-lysine-coated dish. The resulting cell colonies have both pluripotent and stem cell potent characteristics. After culture of cells in the presence of growth factors, cells are differentiated into spermatogonia.

Research Products

• [Journal Article] Effects of extracellular matrices and lectin Dolichos biflorus agglutinin on cell adhesion and self-renewal of bovine gonocytes cultured in vitro (2013)
  • 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
  • Peer Reviewed
• [Journal Article] Protein Kinase A Determines Timing of Early Differentiation through Epigenetic Regulation with G9a (2012)
  • Author(s): Yamamizu K, Fujihara M, Tachibana M, Katayama S, Takahashi A, Hara E, Imai H, Shinkai Y, and Yamashita JK
  • Journal Title: Cell Stem Cell Volume: 10 Issue: 6 Pages: 759-770
  • DOI: 10.1016/j.stem.2012.02.022
  • Peer Reviewed
• [Journal Article] 哺乳動物におけるiPS細胞 (2012)
  • Author(s): 浅野良太
  • Journal Title: 日本胚移植研究会誌 Volume: 34 Pages: 9-18
  • 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. and Imai H
  • Journal Title: Genes to Cells Volume: 16 Issue: 7 Pages: 815-825
  • DOI: 10.1111/j.1365-2443.2011.01528.x
  • NAID: 120005593680
  • Peer Reviewed
• [Journal Article] Characterization and in vitro culture of male germ cells from developing bovine testes (2011)
  • Author(s): Fujihara M, Kim SM, Minami N, Yamada M and Imai H
  • Journal Title: Journal of Reproduction and Development Volume: 57 Pages: 355-364
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/21289464
  • Peer Reviewed
• [Journal Article] Identification of reprogramming factors associated with a pluripotent potential of porcine and bovine spermatic stem cells in culture (2011)
  • Author(s): Kim SM, Fujihara M, Goel S, Sahare M, Minami N, Yamada M and Imai H
  • Journal Title: The Proceedings for 16th World Congress on In Vitro Fertilization Pages: 41-48
  • URL: http://isivf2011.org/jp/
  • Peer Reviewed
• [Journal Article] UCHL-1 protein expression specifically marks spermatogonia in wild bovid testis (2011)
  • Author(s): Goel S, Mahla RS, Suman SK, Reddy N and Imai H
  • Journal Title: European Journal of Wildllife Research Volume: 57 Issue: 3 Pages: 663-667
  • DOI: 10.1007/s10344-010-0454-1
• [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, N., 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 Issue: 17 Pages: 7040-7045
  • DOI: 10.1073/pnas.1013634108
  • Peer Reviewed
• [Journal Article] Pluripotent stem cells from testis (2011)
  • Author(s): Goel S., Imai H.
  • Journal Title: Embryonic Stem Cells-Differentiation and Pluripotent Alternatives(ed.Michael S.Kallos, InTech Publisher, Rijeka) Pages: 473-492
  • Peer Reviewed
• [Journal Article] Identification of reprogramming factors associated with a pluripotent potential of porcine and bovine spermatic stem cells in culture (2011)
  • Author(s): Kim SM, Fujihara M, Goel S, Sahare M, Minami N, Yamada M, Imai H
  • Journal Title: The Proceedings for 16th World Congress on In Vitro Fertilization Pages: 41-48
  • 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 H.
  • Journal Title: Proceedings of the National Academy of Science USA Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] 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)
  • Author(s): Tsukiyama T, Asano R, Kawaguchi T, Kim N, Yamada M, Minami N, Ohinata Y, Imai H.
  • Journal Title: Genes to Cells Volume: (印刷中)
  • NAID: 120005593680
  • Peer Reviewed
• [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: Journal of Reproduction and Development Volume: (印刷中)
  • NAID: 10029050501
  • Peer Reviewed
• [Journal Article] UCHL-1 protein expression specifically marks spermatogonia in wild bovid testis. (2011)
  • Author(s): Goel S, Mahla RS, Suman SK, Reddy N, Imai H.
  • Journal Title: European Journal of Wild Animal Research Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] Spermatogonia-specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia (2010)
  • Author(s): Goel S, Reddy N, Mandal S, Fujihara M, Kim SM and Imai H
  • Journal Title: Theriogenology Volume: 74 Issue: 7 Pages: 1221-1232
  • DOI: 10.1016/j.theriogenology.2010.05.025
  • Peer Reviewed
• [Journal Article] Spermatogonia specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia. (2010)
  • Author(s): Goel S., Reddy N., Mandal S., Fujihara M., Kim SM., Imai H.
  • Journal Title: Theriogenology Volume: 74 Pages: 1221-1232
  • Peer Reviewed
• [Presentation] Long term self-renewal and proliferation of bovine male germ stem cells in vitro (2013)
  • Author(s): Sahare M, Kim SM, Otomo A, Komatsu K, Minami N, Yamada M and Imai H
  • Organizer: CiRA International Symposium 2013 ~Raising the next generation of stem cell research
  • Place of Presentation: Kyoto
• [Presentation] Long term self-renewal and proliferation of bovine male germ stem cells in vitro. (2013)
  • Author(s): 今井 裕
  • Organizer: CiRA International Symposium 2013 ~Raising the next generation of stem cell research~
  • Place of Presentation: Kyoto, Japan
• [Presentation] Mitogen activated protein kinase (MAPK) signaling is indispensable for maintaning self renewal and proliferation of bovine male germ stem cells in culture (2013)
  • Author(s): 今井 裕
  • Organizer: The 2013 Society for Study of Reproduction (SSR) Annual Meeting
  • Place of Presentation: Montreal, Canada
• [Presentation] Glycan epitopes involve in germ cell differentiation during early development of neonatal mice testis (2012)
  • Author(s): Kim SM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • Organizer: The 4th Congress of the Asia Pacific Initiative on Reproduction
  • Place of Presentation: Osaka
• [Presentation] The role of GalNAc glycan residues associated adhesion and self-renewal in cultured bovine gonocytes (2012)
  • Author(s): Kim SM, Fujihara M, Sahare M, Minami N, Yamada M and Imai H
  • Organizer: The 17th International Congress on Animal Reproduction (ICAR)
  • Place of Presentation: Vancouver, Canada
• [Presentation] Determination of distinct germ cell commitment by the specific affinity of the lectin DBA during early development of gonocytes in neonatal mice testis (2012)
  • Author(s): KimSM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • Organizer: Symposium on Functional Genomics of Early Livestock Development
  • Place of Presentation: Banff, Canada
• [Presentation] Glycan epitopes involve in germ cell differentiation during early development of neonatal mice testis. (2012)
  • Author(s): 今井 裕
  • Organizer: The 4th Congress of the Asia Pacific Initiative on Reproduction
  • Place of Presentation: Osaka, Japan
• [Presentation] The role of GalNAc glycan residues associated adhesion and self-renewal in cultured bovine gonocytes. (2012)
  • Author(s): 今井 裕
  • Organizer: The 17th International Congress on Animal Reproduction (ICAR).
  • Place of Presentation: Vancouver, Canada
• [Presentation] Determination of distinct germ cell commitment by the specific affinity of the lectin DBA during early development of gonocytes in neonatal mice testis. (2012)
  • Author(s): 今井 裕
  • Organizer: Symposium on Functional Genomics of Early Livestock Development
  • Place of Presentation: Banff, Canada
• [Presentation] 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)
  • Author(s): 築山智之・今井 裕
  • Organizer: 日本生化学会九州支部例会シンポジウム(招待講演)
  • Place of Presentation: 鹿児島市
  • Year and Date: 2010-05-23
• [Presentation] 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)
  • Author(s): 築山智之・今井裕
  • Organizer: 日本生化学会九州支部例会シンポジウム
  • Place of Presentation: 鹿児島大学(鹿児島)(招待講演)
  • Year and Date: 2010-05-23
• [Presentation] Comparative Changes on Stem Cell Potential of Bovine Male Germ Cells During Testicular Development from Neonatal to Adult (2010)
  • Author(s): Fujihara M, Kim SM, Minami N, Yamada M and Imai H
  • Organizer: The 43rd Annual Meeting of the Study of the Society for Reproduction (SSR)
  • Place of Presentation: Milwaukee USA
• [Presentation] Stem cells of the male germline: an aid to conservation of endangered species (2010)
  • Author(s): Goel S, Mandal S, Reddy N, Mahara R, Suman S, Raj TA, Fujihara M, Kim SM and Imai H
  • Organizer: The 11th International Symposium on Spermatology (ISS)
  • Place of Presentation: Okinawa, Japan
• [Presentation] Lectin DBA specifically recognizes glycan cell surface epitope of gonocytes in developing mouse testis (2010)
  • Author(s): Kim SM, Fujihara M, Goel S, Minami N, Yamada M and Imai H
  • Organizer: The 11th International Symposium on Spermatology (ISS)
  • Place of Presentation: Okinawa, Japan
• [Presentation] Analysis of culture conditions for generation of germ-line competent induced pluripotent stem (iPS) cells in various mammalian species (2010)
  • Author(s): Tsukiyama T, Asano R, Minami N, Yamada M and Imai H
  • Organizer: The 8th Annual Meeting of the International Society for Stem Cell Research
  • Place of Presentation: San Francisco USA
• [Presentation] Comparative Changes on Stem Cell Potential of Bovine Male Germ Cells During Testicular Development from Neonatal to Adult (2010)
  • Author(s): Fujihara M, Kim SM, Minami N, Yamada M, Imai H.
  • Organizer: The 43rd Annual Meeting of the Study of the Society for Reproduction (SSR)
  • Place of Presentation: USA(Milwaukee)
• [Presentation] Stem cells of the male germline : an aid to conservation of endangered species. (2010)
  • Author(s): Goel S, Mandal S, Reddy N, Mahara R, Suman S, Raj TA, Fujihara M, Kim SM, Imai H.
  • Organizer: The 11th International Symposium on Spermatology(ISS)
  • Place of Presentation: 沖縄コンベンションセンター
• [Presentation] Lectin DBA specifically recognizes glycan cell surface epitope of gonocytes in developing mouse testis. (2010)
  • Author(s): Kim SM, Fujihara M, Goel S, Minami N, Yamada M, Imai H.
  • Organizer: The 11th International Symposium on Spermatology(ISS)
  • Place of Presentation: 沖縄コンベンションセンター
• [Presentation] Analysis of culture conditions for generation of germ-line competent induced pluripotent stem(iPS)cells in various mammalian species. (2010)
  • Author(s): Tsukiyama T, Asano R, Minami N, Yamada M, Imai H.
  • Organizer: The 8^<th> Annual Meeting of the International Society for Stem Cell Research
  • Place of Presentation: USA(San Francisco)
• [Book] Pluripotent stem cells from testis."Embryonic Stem Cells - Differentiation and Pluripotent Alternatives" (2011)
  • Author(s): Goel S. and Imai H
  • Publisher: ed. Michael S. Kallos、InTech Publisher、Rijeka
• [Remarks]
  • URL: http://www.reprod.kais.kyoto-u.ac.jp/index.htm
• [Remarks] 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/
• [Remarks]
  • URL: http://www.reprod.kais.kyoto-u.ac.jp/index.htm