May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
The Fate of Trans-Amplifying Cells Analyzed by in vivo Rabbit Model With Stainless Ring Transplantation
Author Affiliations & Notes
  • T. Kawakita
    Keio Univ Sch of Medicine, Shinjuku, Japan
    Ophthalmology, Ophthalmology,
    Tokyo Dental College, Ichikawa, Japan
  • S. Shimmura
    Keio Univ Sch of Medicine, Shinjuku, Japan
    Ophthalmology, Ophthalmology,
    Tokyo Dental College, Ichikawa, Japan
  • K. Higa
    Ophthalmology, Ophthalmology,
    Tokyo Dental College, Ichikawa, Japan
  • J. Shimazaki
    Keio Univ Sch of Medicine, Shinjuku, Japan
    Ophthalmology, Ophthalmology,
    Tokyo Dental College, Ichikawa, Japan
  • K. Tsubota
    Keio Univ Sch of Medicine, Shinjuku, Japan
    Ophthalmology, Ophthalmology,
    Tokyo Dental College, Ichikawa, Japan
  • Footnotes
    Commercial Relationships  T. Kawakita, None; S. Shimmura, None; K. Higa, None; J. Shimazaki, None; K. Tsubota, None.
  • Footnotes
    Support  Grant-in-Aid for Young Scientists (B) (18790131), Grant-in-Aid for Scientific Research (H18-tissue engineering-young -002)
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 6064. doi:https://doi.org/
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      T. Kawakita, S. Shimmura, K. Higa, J. Shimazaki, K. Tsubota; The Fate of Trans-Amplifying Cells Analyzed by in vivo Rabbit Model With Stainless Ring Transplantation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6064. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To investigate the fate of trans-amplifying epithelial cells in the central cornea using a rabbit persistent corneal epithelial defect model created by transplanting a stainless steel ring in the cornea. Using this model, epithelial cells outside the ring failed to migrate inside the ring.

Methods: : A stainless steel ring (8 mm diameter, 300µm wide and 250µm depth) was transplanted into rabbit corneal stroma using 10-0 nylon interrupted sutures. Subsequently we observed wound healing of epithelial defects created within the ring (4, 5, 6, 8mm diameter) compared with control corneas without ring transplants. After 1 week, hematoxylin staining and phenotype analysis was performed by immunostaining for p63, Ki67, and CK3 in a set of animals. Others were continuously observed with repeated epithelial defect of same diameter until cells exhausted.

Results: : Corneal stroma with ring transplantation for 1week showed little inflammation. When epithelium was totally removed inside of the ring, the epithelial defect persisted for at least a month. (n=3) Immunostaining showed similar expression of p63, Ki67, and CK3 compared to normal control. Wound healing was observed in smaller defects (4, 5, 6 mm), however, once cells failed to cover the surface after repeated epithelial removal, cells exhausted on the next day in all cases.

Conclusions: : Trans-amplifying cells may be maintained for more than 1 month when separated from limbal stem cells in vivo. Their fate may depend on replicative senescence, as well as the area required to heal.

Keywords: wound healing • cornea: basic science • cornea: epithelium 
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