Fate of corneal epithelial cells separated from limbus in vivo

Tetsuya Kawakita, Kazunari Higa, Shigeto Shimmura, Machiko Tomita, Kazuo Tsubota, Jun Shimazaki

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Purpose. To characterize corneal epithelial cells separated from limbus in vivo by transplantation of a stainless steel ring with or without creating a defect inside the ring. Methods. A stainless steel ring (diameter, 8 mm; width, 300 μm; depth, 250 μm) was transplanted into rabbit corneal stroma using 10-0 nylon interrupted suture after cutting to a 250 μm depth by corneal vacuum trephine (diameter, 8.0 mm). Epithelial cells were removed inside the ring, and re-epithelization was evaluated after 1 week. Hematoxylin staining and immunostaining against p63, Ki67, and cytokeratin 3 were performed for phenotypic analysis of corneal epithelia. A corneal epithelial defect was centrally created inside the ring (4, 5, and 6 mm diameter) after transplantation. When re-epithelization was achieved, a central epithelial defect was continuously created until cells were exhausted within the ring. The number of created defects was also analyzed to assess the potential of re-epithelialization. Results. Ring-transplanted corneal stroma showed few signs of inflammation, and when epithelium was totally removed from inside the ring, complete epithelial defects were persistent for ≥ 1 month. Corneal sensation was significantly decreased in corneas with the ring (P < 0.05). Immunostaining demonstrated similar expression patterns for p63, Ki67, and cytokeratin3 as the controls. When rings were transplanted into the intact cornea, inside epithelia prevented epithelial defects in vivo for ≤ 6 months. Conclusions. Transient-amplifying cells might maintain homeostasis for >1 month when separated from their limbus in vivo. This model will be useful for future stem cell research or wound healing models.

Original languageEnglish
Pages (from-to)8132-8137
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number11
DOIs
Publication statusPublished - 2011 Oct

Fingerprint

Corneal Stroma
Stainless Steel
Keratin-3
Transplantation
Epithelial Cells
Re-Epithelialization
Stem Cell Research
Corneal Epithelium
Nylons
Hematoxylin
Vacuum
Wound Healing
Cornea
Sutures
Epithelium
Staining and Labeling
Rabbits
Inflammation

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Fate of corneal epithelial cells separated from limbus in vivo. / Kawakita, Tetsuya; Higa, Kazunari; Shimmura, Shigeto; Tomita, Machiko; Tsubota, Kazuo; Shimazaki, Jun.

In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 11, 10.2011, p. 8132-8137.

Research output: Contribution to journalArticle

Kawakita, Tetsuya ; Higa, Kazunari ; Shimmura, Shigeto ; Tomita, Machiko ; Tsubota, Kazuo ; Shimazaki, Jun. / Fate of corneal epithelial cells separated from limbus in vivo. In: Investigative Ophthalmology and Visual Science. 2011 ; Vol. 52, No. 11. pp. 8132-8137.
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