Live-cell Imaging Analysis of Mouse Embryonic Stem Cell Derivation Processes

• Project/Area Number: 23870017
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Developmental biology
• Research Institution: Osaka University
• Principal Investigator: UEDA Jun 大阪大学, 微生物病研究所, 特任助教 (80450394)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: ES 細胞 / ライブセルイメージング / エピジェネティクス / ES細胞 / ES細

Research Abstract

This project aims to identify the nature and origin of ES cells. More specifically, we wanted to know how ES cells are derived from inner cell mass cells (the events that take place during ES cell derivation), and what kind of cells they originated from. To address these questions, we decided to generate multiple reporter mice and carried out live-cell imaging analysis. One of the reporter mice which we generated can visualize DNA methylation status in living condition. This reporter mouse was termed MethylRO(methylation probe in ROSA26locus) and deposited to RIKEN BRC. Using MethylRO, we discovered that DNA methylation pattern undergoes dynamic changes during early embryonic development, whereas DNA methylation pattern was fairly static in ES cells. We are now preparing to carry out live-cell imaging analysis of ES cell derivation process using this MethylROmouse embryo (Ueda et al.In preparation).

Research Products

• [Journal Article] A Genetic and Developmental Pathway from STAT3 to the OCT4-NANOG Circuit is Essential for Maintenance of ICM Lineages in vivo. (2013)
  • Author(s): Do, D.V., Ueda, J., Messerschmidt, D.M., Lorthongpanich, C., Zhou, Y., Feng, B., Guo, G., Lin, P.J., Hossain, Z., Zhang, W., Moh, A., Wu, Q., Robson, P., Ng, H.H., Poellinger, L., Knowles, B.B., Solter, D. and Fu, X.Y.
  • Journal Title: Genes & Dev Volume: (in press)
  • Peer Reviewed
• [Journal Article] Long-term Live-cell Imaging of Mammalian Preimplantation Development and Derivation Process of Pluripotent Stem Cells from the Embryos. (2013)
  • Author(s): Yamagata, K. and Ueda, J.
  • Journal Title: Dev. Growth Differ Volume: 55 Issue: 4 Pages: 378-389
  • DOI: 10.1111/dgd.12048
  • NAID: 40019734964
  • Peer Reviewed
• [Journal Article] PRDM14 ensures naive pluripotency through dual regulation of signaling and epigenetic pathways in mouse embryonic stem cells (2013)
  • Author(s): Yamaji, M., Ueda, J., Hayashi, K., Ohta, H., Yabuta, Y., Kurimoto, K., Nakato, R., Shirahige, K., and Saitou, M
  • Journal Title: Cell Stem Cell Volume: 12 Issue: 3 Pages: 368-382
  • DOI: 10.1016/j.stem.2012.12.012
  • Peer Reviewed
• [Journal Article] Dual Inhibition of Src and GSK3 Maintains Mouse Embryonic Stem Cells, Whose Differentiation Is Mechanically Regulated by Src Signaling. (2012)
  • Author(s): Shimizu, T., Ueda, J., Ho, J.C., Iwasaki, K., Poellinger, L., Harada, I., and Sawada, Y.
  • Journal Title: Stem Cells Volume: 30 Issue: 7 Pages: 1394-1404
  • DOI: 10.1002/stem.1119
  • Peer Reviewed
• [Journal Article] Dual Inhibition of Src and GSK3 Maintains Mouse Embryonic Stem Cells, Whose Differentiation Is Mechanically Regulated by Src Signaling (2012)
  • Author(s): Takeshi Shimizu, et al
  • Journal Title: Stem Cells Volume: (印刷中)
  • Peer Reviewed
• [Presentation] Role of the Hypoxia-Inducible Histone H3K9 Demethylase Jmjd1a and the H3K9 Methyltransferase G9a in Stem Cell Self-Renewal and Tumori genesis (2012)
  • Author(s): Jun Ueda
  • Organizer: Keystone Symposia "Advances in Hypoxic Signaling"
  • Place of Presentation: Banff, Alberta, Canada
  • Year and Date: 2012-02-15
• [Presentation] Hypoxia-inducible Epigentic Regulators Jmjd1a and G9a Antagonistically Control Angiogenesis and Immature Tumorigenic Cell Populations (2012)
  • Author(s): Ueda, J., Lee, K.L., Ho, J.C., Kitajima, S., Yang, H., Sun, W., Fukuhara, N., Zaiden, N., Chan, S.L., Tachibana, M., Shinkai, Y., Kato, H., and Poellinger, L.
  • Organizer: 第10回がんとハイポキシア研究会
  • Place of Presentation: 横浜
• [Presentation] Visualization of DNA Methylation Status in Living Mice (2012)
  • Author(s): Ueda, J., Yamagata, K., Ikawa, M., and Okabe, M.
  • Organizer: 新学術領域研究「哺乳類初期発生の細胞コミュニティー」第5回班会議
  • Place of Presentation: 逗子
• [Presentation] Hypoxia-inducible Epigentic Regulators Jmjd1a and G9a Antagonistically Control Angiogenesis and Immature Tumorigenic Cell Populations. (2012)
  • Author(s): Ho, J.C., Lee, K.L., Ueda, J., Kitajima, S., Yang, H., Sun, W., Fukuhara, N., Zaiden, N., Chan, S.L., Tachibana, M., Shinkai, Y., Kato, H., and Poellinger, L.
  • Organizer: Frontiers in Cancer Science 2012
  • Place of Presentation: NUS, Singapore
• [Presentation] PRDM14 Controls Sensitivity to Extrinsic Signals and Ensures a Naive Epigenome in Mouse Embryonic Stem Cells. (2012)
  • Author(s): Yamaji, M., Ueda, J., Hayashi, K., Ohta, H., Yabuta, Y., Kurimoto, K., Nakato, R., Shirahige, K., and Saitou, M.
  • Organizer: Cold Spring Harbor Meeting "Germ Cells"
  • Place of Presentation: Cold Spring Harbor, NY, USA
• [Presentation] A genetically encoded probe for monitoring the endogenous histone H3 Lys9 acetylation in living cells (2012)
  • Author(s): Sato, Y., Ueda, J., Horikoshi, N., Kujirai, T., Stasevich, T.J., Yamagata, K., Kurumizaka, H., Nozaki, N., and Kimura, H.
  • Organizer: Cold Spring Harbor Meeting "Epigenetics & Chromatin"
  • Place of Presentation: Cold Spring Harbor, NY, USA
• [Presentation] ES 細胞のできるまで (2012)
  • Author(s): 山縣一夫,上田潤,水谷英二,斎藤通紀,若山照彦
  • Organizer: 第105回日本繁殖生物学会大会
  • Place of Presentation: 筑波
• [Presentation] Visualization of DNA Methylation Status in Living Mice (2012)
  • Author(s): Ueda, J., Yamagata, K., Ikawa, M., and Okabe, M.
  • Organizer: 生殖サイクル若手勉強会
  • Place of Presentation: 仙台
• [Presentation] The STAT3-OCT4 Axis is Essential for Development of Inner Cell Mass and Embryonic Stem Cells (2012)
  • Author(s): Do, D.V., Ueda, J., Lorthongpanich, C., Zhang, W., Moh, A., Zhou, Y., Feng, B., Guo, G., Lin, P.J., Hossain, Z., Messerschmidt, D.M., Wu, Q., Robson, P., Ng, H.H., Poellinger, L., Knowles, B.B., Solter, D. and Fu, X.Y.
  • Organizer: International Society for Stem Cell Research, 10th Annual Meeting
  • Place of Presentation: Yokohama, Japan
• [Presentation] ヒストン修飾抗体 scFv を用いた H3K9 アセチル化レベルの生細胞イメージング (2012)
  • Author(s): 佐藤優子,上田潤,山縣一夫,堀越直樹,胡桃坂仁志,野崎直仁,木村宏
  • Organizer: 第6回エピジェネティクス研究会年会
  • Place of Presentation: 東京
• [Presentation] Role of the Hypoxia-Inducible Histone H3K9 Demethylase Jmjd1a and the H3K9 Methyltransferase G9a in Stem Cell Self-Renewal and Tumorigenesis (2012)
  • Author(s): Ueda, J., Lee, K.L., Ho, J.C., Yang, H., Fukuhara, N., Chan, S.L., Tachibana, M., Shinkai, Y., Kato, H., and Poellinger, L.
  • Organizer: Keystone Symposia "Advances in Hypoxic Signaling"
  • Place of Presentation: Banff, Alberta, Canada
• [Presentation] Survival and death of epiblast cells in ES derivation process revealed by long-term live-cell imaging of Oct4 reporter system. (2012)
  • Author(s): Ueda, J., Yamagata, K., Mizutani, E., Saitou, M., and Wakayama, T.
  • Organizer: 定量生物学の会,第四回年会
  • Place of Presentation: 名古屋
• [Presentation] Hypoxia-inducible Epigentic Regulators Jmjd1a and G9a Antagonistically Control Angiogenesis and Immature Tumorigenic Cell Populations. (2012)
  • Author(s): Ueda, J., Lee, K.L., Ho, J.C., Kitajima, S., Yang, H., Sun, W., Fukuhara, N., Zaiden, N., Chan, S.L., Tachibana, M., Shinkai, Y., Kato, H., and Poellinger, L.
  • Organizer: 第10回がんとハイポキシア研究会
  • Place of Presentation: 横浜
• [Book] 細胞 (2013)
  • Author(s): 上田潤,三村維真理
• [Remarks]
  • URL: http://researchmap.jp/wiz/
• [Remarks]
  • URL: http://www.cgabr.biken.osaka-u.ac.jp/
• [Patent(Industrial Property Rights)] Method to preserve the pluripotency of embryonic stem cells. (2011)
  • Inventor(s): Sawada, Y., Shimizu, T., and Ueda, J.
  • Industrial Property Rights Holder: Sawada, Y., Shimizu, T., and Ueda, J.
  • Filing Date: 2011-06-09
  • Overseas
• [Patent(Industrial Property Rights)] Method to preserve the pluripotency of embryonic stem cells (2011)
  • Inventor(s): Yasuhiro Sawada, Takeshi Shimizu, Jun Ueda
  • Industrial Property Rights Holder: Yasuhiro Sawada, Takeshi Shimizu, Jun Ueda
  • Filing Date: 2011
  • Overseas