May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Development of Hybrid–Typed Cultivated Epithelial Sheet Transplantation for Ocular Surface Reconstruction
Author Affiliations & Notes
  • T. Nakamura
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
    Research Center for Regenertive Medicine, Doshisha University, Kyoto, Japan
  • H. Rigby
    Institute of Environmental and Natural Sciences, Lancaster University, Lancaster, United Kingdom
  • E. Sekiyama
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • H. Sogabe
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • N.J. Fullwood
    Institute of Environmental and Natural Sciences, Lancaster University, Lancaster, United Kingdom
  • S. Kinoshita
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Footnotes
    Commercial Relationships  T. Nakamura, None; H. Rigby, None; E. Sekiyama, None; H. Sogabe, None; N.J. Fullwood, None; S. Kinoshita, None.
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 3936. doi:
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      T. Nakamura, H. Rigby, E. Sekiyama, H. Sogabe, N.J. Fullwood, S. Kinoshita; Development of Hybrid–Typed Cultivated Epithelial Sheet Transplantation for Ocular Surface Reconstruction . Invest. Ophthalmol. Vis. Sci. 2006;47(13):3936.

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

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Abstract

Purpose: : To investigate the possibility of using hybrid–typed (including both corneal and oral epithelium) cultivated epithelial sheet transplantation in ocular surface reconstruction.

Methods: : Corneal and oral epithelial cells from adult albino rabbits were isolated, mixed, and cultivated for 2 weeks on a denuded AM carrier (N=4). The resultant hybrid–typed cultivated epithelium was examined by electron microscopy (EM) and immunohistochemically labeled for several keratins. At 2 weeks after the ocular surface injury, the conjunctivalized corneal surfaces of the rabbits were surgically reconstructed by transplanting the hybrid–typed cultivated epithelial sheet.

Results: : The hybrid–typed cultivated epithelial sheet had 4–6 layers of stratified, well differentiated cells. EM examination revealed that the epithelial cells were very similar in appearance to those of normal corneal epithelium. EM revealed that the hybrid–typed sheets were differentiated into columnar, wing, and squamous cells, had numerous desmosmal junctions, and were attached to a basement membrane with hemi–desmosomes. Immunohistochemistry confirmed the presence of keratins 4/13 and 3/12 in the hybrid–typed cultivated epithelial cells. Corneas that were grafted with the hybrid–typed sheets were clear, and were all epithelialized 1 month after surgery.

Conclusions: : We have successfully generated on AM confluent cultures of hybrid–typed epithelial cells expanded ex vivo from biopsy–derived cornea and oral mucosal tissues for ocular surface reconstruction. Our morphological and biological findings indicate that hybrid–typed cultivated epithelial sheets could be useful for ocular surface reconstruction.

Keywords: cornea: epithelium • cornea: basic science • cornea: clinical science 
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