title: Fate of corneal epithelial cells separated from limbus in vivo.
abstract: OBJECTIVE
  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.
authors: Department of Ophthalmology, Tokyo Dental College, Chiba, Japan. kawakita@a2.keio.jp
authors: Tetsuya
authors: T
authors: Kawakita
authors: Kazunari
authors: K
authors: Higa
authors: Shigeto
authors: S
authors: Shimmura
authors: Machiko
authors: M
authors: Tomita
authors: Kazuo
authors: K
authors: Tsubota
authors: Jun
authors: J
authors: Shimazaki
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006706:Homeostasis / Q000502:physiology; D053558:Keratin-3 / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D019736:Prostheses and Implants; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology*
chemical_list: D053558:Keratin-3; D019394:Ki-67 Antigen; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins