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Realization of corneal regenerative medicine using cultured human corneal endothelial and stromal cells

Research Project

Project/Area Number 18390464
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Ophthalmology
Research InstitutionThe University of Tokyo

Principal Investigator

AMANO Shiro  The University of Tokyo, 医学部・附属病院, 准教授 (80193027)

Co-Investigator(Kenkyū-buntansha) 山上 聰 (山上 聡)  東京大学, 医学部附属病院, 准教授 (10220245)
臼井 智彦  東京大学, 医学部附属病院, 助教 (80282557)
杉崎 顕史  東京大学, 医学部附属病院, 助教 (40361480)
Co-Investigator(Renkei-kenkyūsha) YAMAGAMI Satoru  東京大学, 医学部・附属病院, 准教授 (10220245)
USUI Tomohiko  東京大学, 医学部・附属病院, 助教 (80282557)
SUGISAKI Akifumi  東京大学, 医学部・附属病院, 助教 (40361480)
Project Period (FY) 2006 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥2,220,000)
Fiscal Year 2008: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2007: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2006: ¥7,800,000 (Direct Cost: ¥7,800,000)
Keywords角膜 / 再生医療 / 角膜内皮細胞 / 角膜実質細胞
Research Abstract

40歳以上のドナーから得た角膜内皮細胞においては、高率に染色体異常が発生することがわかった。従ってドナーとしては、40歳以下のできるだけ若いドナーが、望ましいことが示唆された。カルチャーインサートの底からトリプシンEDTAを作用させることで、培養角膜内皮細胞シートを作成した。この細胞シートでは、細胞間にNa-K ATPasの発現が見られ、その細胞密度は2000/mm^2を超えていた。この内皮細胞シートの兎への移植実験では、。術後1週間にわたって、移植角膜の透明性が維持され、内皮をつけなかったコントロール群よりも有意に角膜厚が薄く保たれた。もう一つの内皮移植方法として、Descemet's strippingendothelial keratoplasy(DSEK)の術式において、培養角膜内皮細胞を応用する方法を検討した結果では、通常のDSEK の移植片と同等の移植片を培養角膜内皮細胞を用いて作成することが可能であり、in vivo にもいても短期的には、角膜の透明性を維持することができた。培養角膜実質細胞およびその前駆細胞を用いた実験では、ゼラチン上において前駆細胞のほうがより多くの細胞外基質の産生が観察され、兎角膜実質内への移植実験では、前駆細胞を播種したゼラチンでは術後1カ月においてより多くの細胞の残存と細胞外基質の産生が観察され、前駆細胞の角膜実質における再生医療への応用が期待される結果を得ることができた。

Report

(4 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • Research Products

    (26 results)

All 2008 2007 2006

All Journal Article (22 results) (of which Peer Reviewed: 20 results) Presentation (2 results) Book (2 results)

  • [Journal Article] Transplantation of a sheet of human corneal endothelial cell in a rabbit model2008

    • Author(s)
      Hitani K, Yokoo S, Honda N, Usui T, Yamagami S, Amano S
    • Journal Title

      Mol Vis 14

      Pages: 1-9

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Isolation and distribution of rabbit keratocyte precursors2008

    • Author(s)
      Mimura T, Amano S, Yokoo S, Uchida S, Usui T, Yamagami S
    • Journal Title

      Mol Vis 14

      Pages: 197-203

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Decellularizing corneal stroma using N_2 gas2008

    • Author(s)
      Amano S, Shimomura N, Yokoo S, Yamagami S, Araki-Sasaki K
    • Journal Title

      Mol Vis 14

      Pages: 878-882

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Karyotype changes in cultured human corneal endothelial cells2008

    • Author(s)
      Miyai T, Maruyama Y, Osakabe Y, Nejima R, Miyata K, Amano S
    • Journal Title

      Mol Vis 14

      Pages: 942-950

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Human corneal epithelial equivalents for ocular surface reconstruction in complete serum free culture system without unknown factors2008

    • Author(s)
      Yokoo S, Yamagami S, Usui T, Amano S, Araie M
    • Journal Title

      Invest Ophthalmol Vis Sci 49

      Pages: 2438-2443

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] A mouse model of allogeneic corneal endothelial cell transplantation2008

    • Author(s)
      Hayashi T, Yamagami S, Tanaka K, Yokoo S, Usui T, Amano S, Mizuki N
    • Journal Title

      Cornea 27

      Pages: 699-705

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] A novel isolation technique of progenitor cells in human corneal epithelium using uncoated dishes2008

    • Author(s)
      Yokoo S, Yamagami S, Shimada T, Sato T, Usui T, Amano S, Araie M, Hamuro J
    • Journal Title

      Stem Cells 26

      Pages: 1743-1748

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Tissue engineering of corneal stroma with rabbit fibroblast precursors and gelatin hydrogels2008

    • Author(s)
      Mimura T, Amano S, Yokoo S, Uchida S, Yamagami S, Usui T, Kimura Y, Tabata Y
    • Journal Title

      Mol Vis 4

      Pages: 1819-28

    • NAID

      120001851020

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Transplantation of a sheet of human corneal endothelial cell in a rabbit model.2008

    • Author(s)
      Hitani K, Yokoo S, Honda N, Usui T, Yamagami S, Amano S
    • Journal Title

      Molecular Vision 14

      Pages: 1-9

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Isolation and distribution of rabbit keratocyte precursors.2008

    • Author(s)
      Mimura T,Amano S, Yokoo S, Uchida S, Usui T,Yamagami S.
    • Journal Title

      Molecular Vision 14

      Pages: 187-203

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Decellularizing corneal stroma using N2 gas.2008

    • Author(s)
      Amano S,Shimomura N, Yokoo S,Yamagami S, Araki-Sasaki K.
    • Journal Title

      Molecular Vision 14

      Pages: 878-882

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Distribution of precursors in human corneal stromal cells and endothelial cells2007

    • Author(s)
      Yamagami S, Yokoo S, Mimura T, Araie M, Amano S
    • Journal Title

      Ophthalmology 114

      Pages: 433-439

    • Related Report
      2008 Final Research Report 2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] transplantation of conjunctival epithelial cells cultured on amniotic membrane in a rabbit model2007

    • Author(s)
      Ono K, Yokoo S, Mimura T, Usui T, Miyata K, Araie M, Yamagami S, Amano S. Autologous
    • Journal Title

      Mol Vis 13

      Pages: 1138-1143

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Preoperative evaluation of cultured human corneal limbal epithelium on amniotic membrane by confocal microscopy2007

    • Author(s)
      Mimura T, Yamagami S, Usui T, Yokoo S, Ono K, Honda N, Kaneda A, Sugisaki K, Sayegh RR, Amano S
    • Journal Title

      Curr Eye Res 32

      Pages: 407-411

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Necessary prone position time for human corneal endothelial precursor transplantation in a rabbit endothelial deficiency model2007

    • Author(s)
      Mimura T, Yamagami S, Honda N, Usui T, Yokoo S, Amano S
    • Journal Title

      Curr Eye Res 32

      Pages: 617-623

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Characterization of bone marrow-derived cells in the substantia propria of the human conjunctiva2007

    • Author(s)
      Yamagami S, Yokoo S, Amano S, Ebihara N
    • Journal Title

      Invest Ophthalmol Vis Sci 48

      Pages: 4476-4481

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Autologous transplantation of conjunctival epithelial cells cultured on amniotic on amnuotic memvrane in a rabbit model2007

    • Author(s)
      Ono K, Yokoo S, Mimura T, Usui T, Miyata K, Araie M
    • Journal Title

      Molecular Vision 13

      Pages: 1138-1143

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Preoperative evlution of cultured human eorneal limbal epithelium on arnniotic uiembrane by confocal microscopy2007

    • Author(s)
      Mimura T, Yamagami S, Usui T, Yokoo S, Ono K, Honda N, Kaneda A, S
    • Journal Title

      Current Eye Research 32

      Pages: 407-411

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Bone marrow-derived cells in normal human corneal stroma2006

    • Author(s)
      Yamagami S, Ebihara N, Usui T, Yokoo S, Amano S
    • Journal Title

      Arch Ophthalmol 124

      Pages: 62-69

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] UV-absorption in human oral mucosal epithelial sheets for ocular surface reconstruction2006

    • Author(s)
      Yokoo S, Yamagami S, Mimura T, Ono K, Amano S, Saijo H, Mori Y, Takato T
    • Journal Title

      Ophthalmic Res 38

      Pages: 350-354

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Bone marrow-derived cells in normal human corneal stroma.2006

    • Author(s)
      Yamagami S, Ebihara N, Usui T, Yokoo S, Amano S
    • Journal Title

      Archives of Ophthalmology 124

      Pages: 62-69

    • Related Report
      2006 Annual Research Report
  • [Journal Article] UV-absorption in human oral mucosal epithelial sheets for ocular surface reconstruction.2006

    • Author(s)
      Yokoo S, Yamagami S, Mimura T, Ono K, Amano S, Saijo H, Mori Y, Takato T.
    • Journal Title

      Ophthalmic Research 38

      Pages: 350-354

    • Related Report
      2006 Annual Research Report
  • [Presentation] Corneal endothelial cell precursors2008

    • Author(s)
      Amano S, Yamagami S, Usui T, Yokoo S
    • Organizer
      International Congress of Eye Research
    • Place of Presentation
      Beijing, China
    • Related Report
      2008 Final Research Report
  • [Presentation] Karyotype changes in cultured human corneal endothelial cells2008

    • Author(s)
      Amano S, Miyai T, Maruyama Y, Osakabe Y, Nejima R, Miyata K
    • Organizer
      World Ophthalmology Congress
    • Place of Presentation
      Hong Kong, China
    • Related Report
      2008 Final Research Report
  • [Book] 培養角膜内皮細胞移植の現状2008

    • Author(s)
      天野史郎
    • Publisher
      IOL&RS
    • Related Report
      2008 Final Research Report
  • [Book] 角膜内皮における再生医療2007

    • Author(s)
      天野史郎
    • Publisher
      眼科
    • Related Report
      2008 Final Research Report

URL: 

Published: 2006-04-01   Modified: 2016-04-21  

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