Establishment of models for retinal regenerative medicine using the eye-like structures from ES cells

• Project/Area Number: 15390527
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: Gifu University
• Principal Investigator: KUNISADA Takahiro Gifu University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (30205108)
• Co-Investigator(Kenkyū-buntansha): MOTOHASHI Tsutomu Gifu University, Graduate School of Medicine, Research associate, 大学院・医学系研究科, 助手 (40334932) ; INATOMI Tsutomu Kyoto Prefectural University of Medicine, School of Medicine, Research associate, 医学部, 助手 (00305583) ; KINOSHITA Shigeru Kyoto Prefectural University of Medicine, School of Medicine, Professor, 医学部, 教授 (30116024)
• Project Period (FY): 2003 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥16,500,000 (Direct Cost: ¥16,500,000); Fiscal Year 2005: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2004: ¥3,800,000 (Direct Cost: ¥3,800,000); Fiscal Year 2003: ¥9,500,000 (Direct Cost: ¥9,500,000)
• Keywords: ES cells / retina / lens / in vitro differentiation / regenerative medicine / 網膜色素変性症 / 細胞療法 / レンズ / 試験管内分化 / 再生医療

Research Abstract

A culture system to generate eye-like structures consisting of lens, neural retina, and retinal pigmented epithelium (RPE) cells from undifferentiated embryonic stem cells has been established. Precursors of RPE cells that differentiated in the cultures were responsive to Wnt2b signaling and identified retrospectively to form secondary colonies consisting of only RPE-like cells in eye-like structures. These transplanted eye-like structures were capable of populating the developing chick eye as neuronal retina and RPE cells. The outgrowth of a single cell layer of mature RPE cells from the grafted eye-like structures confirmed the existence of precursors for RPE cells. These results suggest that the eye-like structures resulted from the normal developmental pathway responsible for generating eyes in vivo. In addition, eye-like structure-derived cells efficiently regenerated injured retinal ganglion cells treated with NMDA, including those extending axons to retinal papilla. If a functional effect of these cells can be established, such eye-like structures may be potentially used to establish therapy models for various eye diseases.

Research Products

• [Journal Article] Embryonic stem cells that differentiate into RPE cell precursors in vitro develop into RPE cell monolayers in vivo. (2006)
  • Author(s): Aoki, H., Hara, A., Nakagawa, S., Motohashi, T., Hirano, M., Takahashi, Y., Kunisada, T.
  • Journal Title: Exp. Eye Res. 82 Pages: 265-274
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Embryonic stem cells that differentiate into RPE cell precursors in vitro develop into RPE cell monolayers in vivo. (2006)
  • Author(s): Aoki, H., Hara, A., Nakagawa, S., Motohashi, T., Hirano, M., Takahashi, Y., Kunisada, T.
  • Journal Title: Exp.Eye.Res. 82 Pages: 265-274
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Embryonic stem cells that differentiate into RPE cell precursors in vitro develop into RPE cell monolayers in vivo. (2006)
  • Author(s): Aoki, H., Hara, A., Nakagawa, S., Motohashi, T., Hirano, M., Takahashi, Y., Kunisada, T.
  • Journal Title: Exp.Eye Res. 82 Pages: 265-274
• [Journal Article] Cooperative and indispensable roles of endothelin 3 and KIT signalings in melanocyte development. (2005)
  • Author(s): Aoki, H., Motohashi, T., Yoshimura, N., Yamazaki, H., Yamane, T., Panthier, J.J., Kunisada, T
  • Journal Title: Dev. Dyn. 233 Pages: 407-417
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cooperative and indispensable roles of endothelia 3 and KIT signalings in melanocyte development. (2005)
  • Author(s): Aoki, H., Motohashi, T., Yoshimura, N., Yamazaki, H., Yamane, T., Panthier, J.J., Kunisada, T.
  • Journal Title: Dev.Dyn. 233 Pages: 407-417
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cooperative and indispensable roles of endothelin 3 and KIT signalings in melanocyte development. (2005)
  • Author(s): Aoki H., Motohashi, T., Yoshimura, N., Yamazaki, H., Yamane T., Panthier, J.J., Kunisada, T
  • Journal Title: Dev.Dyn. 233 Pages: 407-417
• [Journal Article] Embryonic stem cells are capable of generating a neuronal network in the adult mouse retina.999,216-221,2004. (2004)
  • Author(s): Hara, A., Niwa, M., Kunisada, T., Yoshimura, N., Katayama, M., Kozawa, O., Mori, H.
  • Journal Title: Brain Res. 999 Pages: 216-221
• [Journal Article] 整形外科領域の再生医療(第一回基礎領域) (2004)
  • Author(s): 國貞隆弘
  • Journal Title: 臨床整形外科 39 Pages: 1077-1081
• [Journal Article] A culture method for the induction of neuroshperes from mouse embryonic stem cells by co-culture with PA6 stromal cells
  • Author(s): Kitajima, H., Yoshimura, S., Kokuzawa, J., Kato, M., Iwama, T., Motohashi, T., Kunisada, T., Sakai, N.
  • Journal Title: Mol.Cell Neurosci. In press
• [Book] 培養細胞実験ハンドブック (2004)
  • Author(s): 國貞隆弘
  • Publisher: 羊土社
• [Publications] Hirano, M: "Generarion of structures formed by lens and retinal cells differentiating from embryonic stem cells"Dev.Dyn.. 228. 664-671 (2003)
• [Publications] Hara A.: "Embryonic stem cells are capable of generating a neuronal network in the adult mouse retina"Brain Res.. 999. 216-221 (2004)
• [Publications] Kato, M.: "Hepatocyte growth factor promotes neuronal differentiation of neural stem cells derived from embryonic stem cells"NeuroReport. 15. 5-8 (2004)
• [Publications] 石崎勝彦, 國貞隆弘: "ES細胞からの組織様構造の誘導"Molecular Medicine. 40. 127-127 (2003)
• [Publications] 國貞隆弘: "ES細胞から誘導された眼様構造"再生医療. 2. 59-63 (2003)