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