Single-cell microscopy with stimulus and response device to analyze relaxation mechanism of ES cells fluctuation

• Project/Area Number: 21602001
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Stem cell biology and medical science
• Research Institution: Nagaoka University of Technology (2010-2011); The University of Tokyo (2009)
• Principal Investigator: OHNUMA Kiyoshi 長岡技術科学大学, 産学融合トップランナー養成センター, 特任准教授 (50396834)
• Co-Investigator(Kenkyū-buntansha): WAKAMOTO Yuichi 東京大学, 大学院・総合文化研究科, 准教授 (30517884) ; HEXIG Bayar 東京工業大学, 大学院・生命理工学研究科, 特任助教 (20515168) ; MICHIUE Tatsuo 東京大学, 大学院・総合文化研究科, 准教授 (10282724)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 1細胞培養 / ES細胞 / ゆらぎ / 微細加工 / 初期発生 / 一細胞培養 / 揺らぎ / 無血清培養 / E-カドヘリン

Research Abstract

The mechanism that provides stability during embryonic development remains to be uncovered. To do this, we made a system to observe a single-cell reaction to culture medium stimulation. The system consists of four parts : ES cells, which are a good model for early development, a culture medium without any unknown factors, a specially coated culture dish that enabled us to dissociate cells without aggregation, and time-lapse microscopy. This system enabled us to monitor life-and-death, proliferation, and differentiation of a single ES cell. Single-cell level observation of stimulation response of ES cells will deepen our understanding of developmental stability.

Research Products

• [Journal Article] Glycome diagnosis of human induced pluripotent stem cells using lectin microarray (2011)
  • Author(s): Tateno H, Toyoda M, Saito S, Onuma Y, Ito Y, Hiemori K, Fukumura M, Nakasu A, Nakanishi M, Ohnuma K, Akutsu H, Umezawa A, Horimoto K, Hirabayashi J, Asashima M.
  • Journal Title: J Biol Chem Volume: 286(23) Pages: 20345-20353
  • Peer Reviewed
• [Journal Article] Reduction of N-glycolylneuraminic Acid in Human Induced Pluripotent Stem Cells Generated or cultured under Feeder-and Serum-free Defined Conditions (2010)
  • Author(s): Y. Hayashi, T. Chan, M. Warashina, M. Fukuda, T. Ariizumi, K. Okabayashi, N. Takayama, M. Otsu, K. Eto, M. K. Furue, T. Michiue, K. Ohnuma, H. Nakauchi, M. Asashima
  • Journal Title: PLoS ONE Volume: 5
• [Journal Article] Inhibitory Smad proteins promote the differentiation of mouse embryonic stem cells into ependymal-like ciliated cells (2010)
  • Author(s): Y. Nishimura, A. Kurisaki, M. Nakanishi, K. Ohnuma, N. Ninomiya, S. Komazaki, S. Ishiura, and M. Asashima
  • Journal Title: Biochem Biophys Res Commun Volume: 401(1) Pages: 1-6
• [Journal Article] Induction of neural crest cells from mouse embryonic stem cells in a serum-free monolayer culture (2010)
  • Author(s): Y. Aihara, Y. Hayashi, M. Hirata, N. Ariki, S. Shibata, N. Nagoshi, M. Nakanishi, K. Ohnuma, M. Warashina, T. Michiue, H. Uchiyama, H. Okano, M. Asashima, and M. Kusuda Furue
  • Journal Title: Int J Dev Biol Volume: 54 Pages: 1287-94
• [Journal Article] An in vitro reconstitution system for the assessment of chromatin protein fluidity during Xenopus development (2010)
  • Author(s): R. Aoki, M. Inui, Y. Hayashi, A. Sedohara, K. Okabayashi, K. Ohnuma, M. Murata, and M. Asashima
  • Journal Title: Biochem Biophys Res Commun Volume: 400 Pages: 200-6
• [Journal Article] BMP4 induction of trophoblast from mouse embryonic stem cells in defined culture conditions on laminin (2010)
  • Author(s): Hayashi H, Furue MK, Satoshi Tanaka S, Hirose M, Wakisaka N, Danno H, Ohnuma K, Oeda S, Aihara Y, Shiota K, Ogura A, Ishiura S and Asashima M
  • Journal Title: In Vitro Cell Dev Biol Anim Volume: 46(5) Pages: 416-416
• [Journal Article] Pdx1-transfected adipose tissue-derived stem cells differentiate into insulin-producing cells in vivo and reduce hyperglycemia in diabetic mice (2010)
  • Author(s): Kajiyama H, Hamazaki S T, Tokuhara M, Masui S, Okabayashi K, Ohnuma K, Yabe S, Yasuda K, Ishiura S, Okochi H and Asasima M
  • Journal Title: IJDB Volume: 54(4) Pages: 699-705
• [Journal Article] An in vitro reconstitution system for the assessment of chromatin protein fluidity during Xenopus development.(2010) (2010)
  • Author(s): R.Aoki, 他7名
  • Journal Title: Biochem Biophys Res Commun Volume: 400 Pages: 200-206
  • Peer Reviewed
• [Journal Article] Induction of neural crest cells from mouse embryonic stem cells in a serum-free monolayer culture (2010)
  • Author(s): Y.Aihara, 他13名
  • Journal Title: Int J Dev Biol Volume: 54 Pages: 1287-12940
  • Peer Reviewed
• [Journal Article] Directional Migration of Neuronal PC12 Cells in a Ratchet Wheel-Shaped Microchamber (2009)
  • Author(s): Ohnuma K, Toyota T, Ariizumi T, Sugawara T, Asashima M
  • Journal Title: J Biosci Bioeng Volume: 108 Pages: 76-83
  • NAID: 110007359687
• [Journal Article] Recent progress and perspective of developmental and stem cell biology for regenerative medicine (2009)
  • Author(s): Okabayashi K, Ohnuma K, Asashima M
  • Journal Title: 再生医療 8 Pages: 414-9
  • NAID: 40016914353
• [Journal Article] Development of in vitro differentiation systems using vertebrate stem cells (2009)
  • Author(s): Okabayashi K, Ohnuma K, Asashima M
  • Journal Title: Inflammation and Regeneration 29 Pages: 302-309
  • NAID: 10026094005
• [Journal Article] BMP4 induction of trophoblast from mouse embryonic stem cells in defined culture conditions on laminin
  • Author(s): Hayashi H, Furue MK, Satoshi Tanaka S, Hirose M, Wakisaka N, Danno H, Ohnuma K, Oeda S, Aihara Y, Shiota K, Ogura A, Ishiura S, 〓 M.
  • Journal Title: In Vitro Cell Dev Biol Anim (in press)
  • Peer Reviewed
• [Presentation] ヒトiPS細胞を用いた無血清・無フィーダー培養による心筋誘導系の確立 (2011)
  • Author(s): 小池怜, 伊藤弓弦, 吉満亮介, 丸山篤史, 大沼清
  • Organizer: 日本バイオマテリアル学会
  • Place of Presentation: 京都
  • Year and Date: 2011-11-22
• [Presentation] ヒトiPS細胞培養のためのpolydimethylsiloxaneへの細胞外基質コート条件の探索 (2011)
  • Author(s): 吉満亮介、服部浩二, 杉浦慎治, 金森敏幸, 小池怜, 丸山篤史, 大沼清
  • Organizer: 日本バイオマテリアル学会
  • Place of Presentation: 京都
  • Year and Date: 2011-11-22
• [Presentation] Time-lapse imaging of mES cells monolayer-cultured for the detection of fluctuations between differentiated and undifferentiated states (2011)
  • Author(s): A Maruyama, Y Wakamoto, K Oonuma
  • Organizer: 日本生物物理学会
  • Place of Presentation: 姫路
  • Year and Date: 2011-09-17
• [Presentation] Generation of Human Induced Pluripotent Stem Cells from Adult Dermal Fibroblast under Feeder- and Serum-free Defined Conditions (2011)
  • Author(s): Y.Hayashi, T.Chan, M.Warashina, M.Fukuda, T.Ariizumi, K.Okabayashi, N.Takayama,M.Otsu, K.Eto, M.K.Furue, T.Michiue, K.Ohnuma*(発表者・責任著者), H.Nakauchi, M.Asashima
  • Organizer: 国際幹細胞学会(ISSCR)
  • Place of Presentation: カナダ オンタリオ州
  • Year and Date: 2011-06-16
• [Book] Innovations in Biotechnology, Intech, E. C. Agbo(Ed) (2012)
  • Author(s): T. Toyota, Y. Wakamoto, K. Hayashi and K. Ohnuma
  • Publisher: Controlling Cell Migration with Micropatterns
• [Book] "Controlling Cell Migration with Micropatterns" Innovations in Biotechnology, 2012, ISBN 978-953-51-0096-6, chapter 9 (187-208)(E.C.Agbo (Ed)) (2012)
  • Author(s): T.Toyota, Y.Wakamoto, K.Hayashi, K.Ohnuma
  • Total Pages: 22
  • Publisher: Intech
• [Book] Mechanosensing Biology, 1st Edition., Noda, M.(Ed.) (2011)
  • Author(s): M. Asashima, T. Michiue, K. Ohnuma, Y. Nakajima, Y. Ito
  • Publisher: Mechanobiology During Vertebrate Organ Development
• [Book] "Mechanobiology During Vertebrate Organ Development" Mechanosensing Biology, 1st Edition., 2011, XIV, ISBN : 978-4-431-89756-9, Part I, Chapter 3 (39-50)(Noda, M.(Ed.)) (2011)
  • Author(s): M.Asashima, T.Michiue, K.Ohnuma, Y.Nakajima, Y.Ito
  • Total Pages: 12
  • Publisher: Springer
• [Remarks]
  • URL: http://www.nagaokaut.ac.jp/j/annai/NUT-toprun/ohnuma0.htm