Chemical genomics approach to human embryonic stem cell-self renewal

• Project/Area Number: 22590269
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: KAWASE Eihachiro 京都大学, 再生医科学研究所, 講師 (70402790)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ヒトES細胞 / 化合物ライブラリ / 未分化維持 / 分化制御

Research Abstract

Human embryonic stem cells (hESCs) and induced pluripotent cells have the potential to provide an unlimited source of tissues for regenerative medicine. For this purpose, development of defined/xeno-free culture systems under feeder-free conditions is essential for the expansion of hESCs. Most defined/xeno-free media for the culture of hESCs contain basic fibroblast growth factor (bFGF). Therefore, bFGF is thought to have an almost essential role for the expansion of hESCs in an undifferentiated state. Here, we report identification of small molecules, some of which were neurotransmitter antagonists (trimipramine and ethopropazine), which promote long-term hESC self-renewal without bFGF in the medium. The hESCs maintained high expression levels of pluripotency markers, had a normal karyotype after 20 passages, and could differentiate into all three germ layers.

Research Products

• [Journal Article] Identification of small molecules that promote human embryonic stem cell self-renewal. (2013)
  • Author(s): Kumagai, H., Suemori, H., Uesugi, M., Nakatsuji, N. and Kawase, E
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 434 Issue: 4 Pages: 710-716
  • DOI: 10.1016/j.bbrc.2013.03.061
  • Peer Reviewed
• [Journal Article] Recombinant E8 fragments of human laminin isoforms support long-term culture of human pluripotent stem cells under defined and xeno-free culture conditions. (2012)
  • Author(s): Miyazaki, T., Futaki, S., Suemori, H., Taniguchi,Y., Kawasaki, M., Hayashi, M., Kumagai,H., Nakatsuji, N., Sekiguchi, K. and Kawase, E
  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 1236-1236
  • DOI: 10.1038/ncomms2243
  • Peer Reviewed
• [Journal Article] Efficient and Accurate Homologous Recombination in hESCs and hiPSCs Using Helper-dependent Adenoviral Vectors (2011)
  • Author(s): Aizawa E, Hirabayashi Y, Iwanaga Y, Suzuki K, Sakurai K, Shimoji M, Aiba K, Wada T, Tooi N, Kawase E, Suemori H, Nakatsuji N, Mitani K
  • Journal Title: Mol Ther Volume: 6 Issue: 2 Pages: 424-431
  • DOI: 10.1038/mt.2011.266
  • Peer Reviewed
• [Journal Article] Cardiomyocytes develop from anterior primitive streak cells induced by β-catenin activation and the blockage of BMP signaling in hESCs. (2010)
  • Author(s): Yamauchi, K., Sumi, T., Minami, I., Otsuji, T.G., Kawase, E., Nakatsuji, N., Suemori, H
  • Journal Title: Genes to Cells Volume: 15 (12) Issue: 12 Pages: 1216-1227
  • DOI: 10.1111/j.1365-2443.2010.01455.x
  • Peer Reviewed
• [Journal Article] Role of SOX2 in maintaining pluripotency of human embryonic stem cells (2010)
  • Author(s): Adachi, K., Suemori, H., Yasuda, S. -Y., Nakatsuji N. and Kawase, E
  • Journal Title: Genes to Cells Volume: 15 (5) Pages: 455-469
  • Peer Reviewed
• [Journal Article] Efficient system to integrate functional genes into a defined locus in human embryonic stem cell lines (2010)
  • Author(s): Sakurai, K., Shimoji, M., Tahimic, CGT., Aiba, K., Kawase, E., Hasegawa, K, Amagai, Y., Suemori, H., Nakatsuji, N
  • Journal Title: Nucleic Acids Res Volume: 38 (7)
  • Peer Reviewed
• [Journal Article] Efficient system to integrate functional genes into a defined locus in human embryonic stem cell lines. (2010)
  • Author(s): Sakurai, et al.
  • Journal Title: Nucleic Acids Res. Volume: 38
  • Peer Reviewed
• [Journal Article] Role of SOX2 in maintaining pluripotency of human embryonic stem cells. (2010)
  • Author(s): Adachi, et al.
  • Journal Title: Genes to Cells Volume: 15 Pages: 455-469
  • Peer Reviewed
• [Journal Article] Cardiomyocytes develop from anterior primitive streak cells induced by β-catenin activation and the blockage of BMP signaling in hESCs. (2010)
  • Author(s): Yamauchi, et al.
  • Journal Title: Genes to Cells Volume: 15 Pages: 1216-1227
  • Peer Reviewed
• [Presentation] ラミニンE8フラグメントを用いたヒトES/iPS細胞の単一分散培養法 (2013)
  • Author(s): 宮崎隆道、二木杉子、末盛博文、山田雅司、川崎美和、林麻利亜、熊谷英明、中辻憲夫、関口清俊、川瀬栄八郎
  • Organizer: 第12回日本再生医療学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2013-03-23
• [Presentation] Fluorescent Chemical Probes for Human Stem Cells (2013)
  • Author(s): Nao Hirata, Masato Nakagawa, Yuto Fujibayashi, Kaori Yamauchi, Asako Murata,Itsunari Minami, Maiko Tomioka, Takayuki Kondo, Ting-Fang Kuo, Hiroshi Endo,Haruhisa Inoue, Shin-ichi Sato, Shin Ando,Yoshinori Kawazoe, Kazuhiro Aiba, Koh
  • Organizer: Cira International Symposium
  • Place of Presentation: Kyoto
  • Year and Date: 2013-03-11
• [Presentation] Fluorescent Chemical Probes for Human Stem Cells. (2013)
  • Author(s): Nao Hirata et al.,
  • Organizer: 2013 Cira International Symposium
  • Place of Presentation: Kyoto
• [Presentation] RECOMBINANT HUMAN LAMININ E8 FRAGMENTS (LM-E8s) SUPPORT THE EFFICIENT ADHESION AND EXPANSION OF DISSOCIATED HUMAN PLURIPOTENT STEM CELLS UNDER DEFINED AND XENO-FREE CONDITION (2012)
  • Author(s): Takamichi Miyazaki, Sugiko Futaki, Hirofumi Suemori, Yukimasa Taniguchi, Masashi Yamada, Miwa Kawasaki, Maria Hayashi, Hideaki Kumagai, Norio Nakatsuji, Kiyotoshi Sekiguchi, Eihachiro Kawase
  • Organizer: 2012 World Stem Cell Summit
  • Place of Presentation: West Palm Beach, USA
  • Year and Date: 2012-12-03
• [Presentation] Recombinant E8 fragments of human laminin isoforms support the efficient adhesion and expansion of dissociated human pluripotent stem cells under defined and xeno-free condition. (2012)
  • Author(s): Takamichi Miyazaki, Sugiko Futaki, Hirofumi Suemori, Yukimasa Taniguchi, Masashi Yamada, Miwa Kawasaki, Maria Hayashi, Hideaki Kumagai, Norio Nakatsuji, Kiyotoshi Sekiguchi, Eihachiro Kawase
  • Organizer: 10thInternational Society for Stem Cell Research
  • Place of Presentation: Yokohama
  • Year and Date: 2012-06-15
• [Presentation] Manufacturing and banking of clinical-garade human embryonic stem cell lines in a GMP facility (2012)
  • Author(s): Takada, Kei, Hirai, Masako, Kawase, Eihachiro, Hamao, Mari, Kashigi, Fumi, Suemori, Hirofumi, Nakatsuji, Norio, Takahashi, Tsuneo A
  • Organizer: 10thInternational Society for Stem Cell Research
  • Place of Presentation: Yokohama
  • Year and Date: 2012-06-15
• [Presentation] Applications of scalable culture system for clinical-grade undifferentiated human embryonic stem cells. (2012)
  • Author(s): Kawase E
  • Organizer: Wednesday Industry Symposia, 10thInternational Society for Stem Cell Research
  • Place of Presentation: Yokohama
  • Year and Date: 2012-06-13
• [Presentation] RECOMBINANT HUMAN LAMININ E8 FRAGMENTS (LM-E8s) SUPPORT THE EFFICIENT ADHESION AND EXPANSION OF DISSOCIATED HUMAN PLURIPOTENT STEM CELLS UNDER DEFINED AND XENO-FREE CONDITION. (2012)
  • Author(s): Kawase, E. et al.,
  • Organizer: 2012 World Stem Cell Summit
  • Place of Presentation: West Palm Beach、アメリカ合衆国
• [Presentation] Recombinant E8 fragments of human laminin isoforms support the efficient adhesion and expansion of dissociated human pluripotent stem cells under defined and xeno-free condition. (2012)
  • Author(s): Miyazaki, T. et al.,
  • Organizer: 10th International Society for Stem Cell Research
  • Place of Presentation: Yokohama
• [Presentation] Manufacturing and banking of clinical-garade human embryonic stem cell lines in a GMP facility. (2012)
  • Author(s): Takada, K. et al.,
  • Organizer: 10th International Society for Stem Cell Research
  • Place of Presentation: Yokohama
• [Presentation] Applications of scalable culture system for clinical-grade undifferentiated human embryonic stem cells. (2012)
  • Author(s): Kawase, E.
  • Organizer: Wednesday Industry Symposia, 10th International Society for Stem Cell Research
  • Place of Presentation: Yokohama
  • Invited
• [Presentation] Fluorescent chemical probes for human stem cells (2011)
  • Author(s): Nao Hirata,Masato Nakagawa,Yuto Fujibayashi, Kaori Yamauchi, Asako Murata, Eihachiro Kawase, Shin-ichi Sato, Shin Ando,Young-Tae Chang, Hirofumi Suemori,Norio Nakatsuji,Kazumitsu Ueda, Shinya Yamanaka, Motonari Uesugi.
  • Organizer: AIMECS11 8thAFMC International Medical Chemistry Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2011-11-29
• [Presentation] Fluorescent chemical probes for human stem cells (2011)
  • Author(s): Nao Hirata, et al
  • Organizer: IMECS11 8th AFMC International Medical Chemistry Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2011-11-29
• [Presentation] Homologous Recombination in human pluripotent stem cells using helper-dependent adenovirua vectors (2011)
  • Author(s): Suzuki, K., Hirabayashi, Y., Aizawa, E., Iwanaga, Y., Tokumitsu, A., Sakurai, K., Shimoji, M., Aiba, K., Wada, T., Tooi, N., Kawase, E. Suemori, H., Nakatsuji, N. and Mitani, K
  • Organizer: 9thInternational Society for Stem Cell Research
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2011-06-17
• [Presentation] Homologous Recombination in human pluripotent stem cells using helper-dependent adenovirua vectors (2011)
  • Author(s): Suzuki, K., et al
  • Organizer: 9th International Society for Stem Cell Research
  • Place of Presentation: Toronto(カナダ)
  • Year and Date: 2011-06-17
• [Presentation] ヒトES細胞の未分化維持因子の探索を (2011)
  • Author(s): 熊谷英明、末盛博文、上杉志成、中辻憲夫、川瀬栄八郎
  • Organizer: 第10回日本再生医療学会総会
  • Place of Presentation: 東京
  • Year and Date: 2011-03-01
• [Presentation] ヒトES細胞の未分化維持因子の探索を目的としたHigh content analysis (HCA) (2011)
  • Author(s): 熊谷英明、末盛博文、上杉志成、中辻憲夫、川瀬栄八郎
  • Organizer: 第10回日本再生医療学会総会
  • Place of Presentation: 東京
  • Year and Date: 2011-03-01
• [Presentation] β-cateninの活性化およびBMPシグナルの抑制による原条前側領域の形成を経由したヒトES細胞から心筋細胞への分化 (2010)
  • Author(s): 山内香織、角智行、南一成、尾辻智美、川瀬栄八郎、中辻憲夫、末盛博文
  • Organizer: 第33回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-10
• [Presentation] High Content Analysis (HCA)によるヒトES細胞の未分化維持因子の探索 (2010)
  • Author(s): 熊谷英明、末盛博文、上杉志成、中辻憲夫、川瀬栄八郎
  • Organizer: High Content Analysis (HCA)によるヒトES細胞の未分化維持因子の探索
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-10
• [Presentation] High Content Analysis (HCA)によるヒトES細胞の未分化維持因子の探索 (2010)
  • Author(s): 熊谷英明、末盛博文、上杉志成、中辻憲夫、川瀬栄八郎
  • Organizer: 第33回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-09
• [Presentation] Gene targeting in human pluripotent stem cells with adeno-associated virus vectors (2010)
  • Author(s): Ko Mitani, Kaoru Mitsui, Keiichiro Suzuki, Emi Aizawa, Haruka Shiiba, Eihachiro Kawase, Hirofumi Suemori, Norio Nakatsuji
  • Organizer: 第16回日本遺伝子治療学会
  • Place of Presentation: 栃木
  • Year and Date: 2010-07-01
• [Book] 実験医学4月号 (2013)
  • Author(s): 川瀬栄八郎、宮崎隆道、関口清俊
  • Total Pages: 31
• [Book] 「ラミニンE8を用いた安全で効率的なヒト多能性幹細胞培養法の開発」in 「実験医学」4月号カレントトピックス (2013)
  • Author(s): 川瀬栄八郎
  • Total Pages: 5
  • Publisher: 羊土社
• [Book] Stem Cells in Clinic and Research (2011)
  • Author(s): Ali Gholamrezanezhad
  • Publisher: InTech
• [Book] The role of SOX2 in maintaining pluripotency and differentiation of human embryonic stem "Stem Cells in Clinic and Research" pp.169-184 (Ed.Ali Gholamrezanezhad) (2011)
  • Author(s): Adachi, K., et al
  • Total Pages: 16
  • Publisher: InTech
• [Remarks] 細胞接着タンパクを用い安全・高効率なヒトES/iPS細胞の培養法を開発
  • URL: http://www.icems.kyoto-u.ac.jp/j/pr/2012/12/05-nr.html
• [Remarks] 細胞接着タンパクを用い、 安全・高効率なヒトES/iPS細胞の培養法を開発
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2012/121205_1.htm
• [Remarks] 細胞接着タンパクを用い安全・高効率なヒトES/iPS細胞の培養法を開発
  • URL: http://www.icems.kyoto-u.ac.jp/j/pr/2012/12/05-nr.html
• [Remarks] 細胞接着タンパクを用い、 安全・高効率なヒトES／iPS細胞の培養法を開発
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2012/121205_1.htm