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Establishment of fundamental technology for clinically applicable regenerative medicine in corneal endothelium and stroma.

Research Project

Project/Area Number 21390465
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  東京大学, 医学部附属病院, 教授 (80193027)

Co-Investigator(Kenkyū-buntansha) USUI Tomohiko  東京大学, 医学部附属病院, 講師 (80282557)
SUGISAKI Kenji  東京大学, 医学部附属病院, 助教 (40361480)
Project Period (FY) 2009 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2011: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2010: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2009: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000)
Keywords角膜 / 再生医療 / 角膜内皮 / 角膜実質
Research Abstract

Long-term efficacy of transplantation of cultured human corneal endothelial cells(HCEC) with Descemet's stripping automated endothelial keratoplasty(DSAEK) was investigated. DSAEK grafts with cultured HCEC had normal stromal lamellar structure and endothelial cell layer with intercellular adhesion structure. The corneas in which cultured HCEC were thinner than those in the control group and were transparent at 1 year after the surgery. These results indicated that DSAEK using cultured HCEC is effective and clinically feasible.
The other method of seeding cultured HCEC on the posterior corneal stroma which was obtained after a thick corneal flap was made using a microkeratome, was confirmed to be effective and safe.
A new temperature-responsive culture dish was developed and evaluated. The results indicated that the temperature-responsive culture dish can produce a sheet of corneal epithelium containing a lot of progenitor cells after culture with non-serum and non-feeder cells. We plan to use this new temperature-responsive culture dish to produce a cell sheet of corneal endothelial cells.
A new culture method of HCEC using bi-phosphate ascorbic acid(Asc-2P) was investigated. The results indicated that the primary culture of HCEC was successful in all cases under the culture condition with athello-collagen as a carrier and Asc-2P and bGFGF in the media. Even after several passage, HCEC showed a high proliferative capacity and a cobble-stone like appearance. The expression level and intrecellular distribution pattern of ZO-1 and NA+/K+-ATPase in cultured HCEC were similar to those in in vivo HCEC. 8-OHdG and MDA increased in HCEC cultured without Asc-2P as passage increased whereas they were suppressed in HCEC cultured with Asc-2P. HCEC cultured with Asc-2P showed high proliferative capacity and appearance similar to in vivo HCEC. These results suggested that Asc-2P elongated the lifespan of HCEC by reducing intracellular oxidative stress.

Report

(4 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Research Products

    (25 results)

All 2012 2011 2010 2009

All Journal Article (12 results) (of which Peer Reviewed: 11 results) Presentation (7 results) Book (6 results)

  • [Journal Article] Prospects for Descemet stripping automated endothelial keratoplasty using cultured human corneal endothelial cells2011

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

      J Transplant Technol Res

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Prospects for Descemet stripping automated endothelial keratoplasty using cultured human corneal endothelial cells2011

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

      J Transplant Technol Res

      Volume: S2 Issue: S2 Pages: 1-1

    • DOI

      10.4172/2161-0991.s2-001

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Selective isolation of young cells from human corneal endothelium by the sphere-forming assay2010

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

      Tissue Eng Part C Methods

      Volume: 16 Pages: 803-812

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Isolation of adult progenitor cells with neuronal potential from rabbit corneal epithelial cells in serum-and feeder layer-free culture conditions2010

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

      Mol Vis

      Volume: 16 Pages: 1712-1719

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Biology of corneal endothelial cells in vivo and in vitro2010

    • Author(s)
      Amano S, Kaji Y, Mimura T
    • Journal Title

      Jpn J Ophthalmol

      Volume: 54 Pages: 211-214

    • NAID

      10027204090

    • Related Report
      2011 Final Research Report
  • [Journal Article] Selective isolation of young cells from human corneal endoth elium by the sphere-forming assay.2010

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

      Tissue Eng Part C Methods

      Volume: 116 Pages: 803-812

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Isolation of adult progenitor cells with neuronal potential from rabbit corneal epithelial cells in serum-and feeder layer-free culture conditions.2010

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

      Molecular Vision

      Volume: 16 Pages: 1712-1717

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Effect of latanoprost on the expression of matrix metalloprot einases and tissue inhibitor of metalloproteinase-1 on the ocular surface.2010

    • Author(s)
      Honda N, Miyai T, Nejima R, Miyata K, Mimura T, Usui T, Aihara M, Araie M, Amano S.
    • Journal Title

      Archives of Ophthalmology

      Volume: 128 Pages: 466-471

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Descemet's stripping automated endothelial keratoplasty using cultured corneal endothelial cells in a rabbit model2009

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

      Arch Ophthalmol

      Volume: 127 Pages: 1321-1326

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Immunological mechanisms of corneal allografts reconstituted from cultured allogeneic endothelial cells in an immune-privileged site.2009

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

      Investigative Ophthalmology Visual Science 50

      Pages: 3151-3158

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Descemet's stripping automated endothelial keratoplasty using cultured corneal endothelial cells in a rabbit model.2009

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

      Archives of Ophthalmology 127

      Pages: 1321-1326

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Proposed diagnostic criteria for obstructive meibomian gland dysfunction.2009

    • Author(s)
      Arita R, Itoh K, Maeda S, Maeda K, Furuta A, Fukuoka S, Tomidokoro A, Amano S.
    • Journal Title

      Ophthalmology 116

      Pages: 2058-2063

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Presentation] アスコルビン酸-2リン酸を用いたヒト角膜内皮細胞の培養2012

    • Author(s)
      島伸行、木本美和、山口昌大、天野史郎、山上聡
    • Organizer
      角膜カンファランス2012
    • Place of Presentation
      東京
    • Year and Date
      2012-02-25
    • Related Report
      2011 Annual Research Report 2011 Final Research Report
  • [Presentation] 無血清・無フィーダ-培養に対応した新規温度応答性培養皿による角膜上皮シート2012

    • Author(s)
      横尾誠一、高田哲夫、山上聡、原口和敏、天野史郎
    • Organizer
      角膜カンファランス2012
    • Place of Presentation
      東京
    • Year and Date
      2012-02-25
    • Related Report
      2011 Final Research Report
  • [Presentation] 無血清・無フィーダー培養に対応した新規温度応答性培養皿による角膜上皮シート2012

    • Author(s)
      横尾誠一、高田哲夫、山上聡、原口和敏、天野史郎
    • Organizer
      角膜カンファランス2012
    • Place of Presentation
      東京
    • Year and Date
      2012-02-25
    • Related Report
      2011 Annual Research Report
  • [Presentation] 角膜再生の過去、現在、未来2011

    • Author(s)
      天野史郎
    • Organizer
      第52回日本視能矯正学会特別講演
    • Place of Presentation
      横浜市(招待講演)
    • Year and Date
      2011-11-19
    • Related Report
      2011 Annual Research Report
  • [Presentation] 角膜再生の現在と未来2011

    • Author(s)
      天野史郎
    • Organizer
      第81回九州眼科学会
    • Place of Presentation
      那覇市(招待講演)
    • Year and Date
      2011-05-27
    • Related Report
      2011 Annual Research Report
  • [Presentation] 培養ヒト角膜内皮細胞を用いたウサギのDescemet stripping endothelial keratoplasty2009

    • Author(s)
      本田紀彦、三村達哉、臼井智彦, 天野史郎、新家眞
    • Organizer
      日本眼科学会総会
    • Place of Presentation
      東京国際フォーラム
    • Year and Date
      2009-04-17
    • Related Report
      2009 Annual Research Report
  • [Presentation] An Animal Model of Descemet's Stripping Automated Endothelial Keratoplasty Using Cultured Corneal Endothelial Cells2009

    • Author(s)
      Amano S, Honda N, Usui T, Mimura T
    • Organizer
      ARVO Meeting
    • Place of Presentation
      Fort Lauderdale
    • Year and Date
      2009-04-11
    • Related Report
      2011 Final Research Report
  • [Book] Regenerative Medicine and Tissue Engineering ; From Cells to Organs/Book 22011

    • Author(s)
      Mimura T, Tabata Y, Amano S
    • Related Report
      2011 Final Research Report
  • [Book] Transplantation of corneal stroma reconstructed with gelatin and multipotent precursor cells from corneal stroma. Tissue Engineering for Tissue and Organ Regeneration2011

    • Author(s)
      Mimura T, Tabata Y, Amano S
    • Total Pages
      16
    • Publisher
      Intech
    • Related Report
      2011 Annual Research Report
  • [Book] 角膜内皮細胞はなぜ増えないのか?眼のサイエンス、眼疾患の謎2010

    • Author(s)
      天野史郎
    • Publisher
      文光堂,東京
    • Related Report
      2011 Final Research Report
  • [Book] 正常者の角膜内皮細胞.あたらしい眼科2009

    • Author(s)
      天野史郎
    • Related Report
      2011 Final Research Report
  • [Book] DSAEKの合併症と対策.眼科手術2009

    • Author(s)
      天野史郎
    • Related Report
      2011 Final Research Report
  • [Book] 全層角膜移植術の長期予後.前田直行編集,眼科診療のスキルアップ2009

    • Author(s)
      天野史郎
    • Publisher
      メディカルビュー,東京
    • Related Report
      2011 Final Research Report

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Published: 2009-04-01   Modified: 2016-04-21  

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