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(Investigative Ophthalmology and Visual Science. 2003;44:4698-4704.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.03-0272
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Propagation and Phenotypic Preservation of Rabbit Limbal Epithelial Cells on Amniotic Membrane

Der-Yuan Wang,1,2 Yi-Jen Hsueh,1 Vivian C. Yang,3 and Jan-Kan Chen1

1From the Department of Physiology, College of Medicine, Chang Gung University, Taoyuan, Taiwan; the 2Section of Biologics, Division of Drug Biology, Bureau of Food and Drug Analysis, Department of Health, Executive Yuan, Taipei, Taiwan; and the 3Department of Biology, Tunghai University, Taichung, Taiwan.

PURPOSE. To describe the phenotypic characteristics of a limbal epithelial cell sheet outgrowth from a limbal explant cultured on amniotic membrane.

METHOD. Immunofluorescent staining and confocal microscopy were used to examine the expressions of p63, Ki-67, keratins 3 and 14, connexin 43, and the integrin {alpha}6/ß4 and {alpha}3/ß1 subunits in corneal and limbal tissues in a limbal explant and epithelial outgrowth cultured for 2 weeks on amniotic membrane.

RESULTS. The expression patterns of p63, Ki-67, keratins, integrins, and connexin 43 in a limbal explant with an epithelial outgrowth cultured for 2 weeks on amniotic membrane resembled those in freshly prepared limbus. Moreover, the distribution of integrin subunits in positive cells of the limbal explant and its epithelial outgrowth was similar to that of the corneal epithelial cells during wound repair.

CONCLUSIONS. The epithelial cell sheet grown from a limbal explant on amniotic membrane exhibited a phenotype similar to that of the limbus, suggesting that amniotic membrane is a substrate capable of supporting the propagation and preservation of p63-positive limbal epithelial cells.




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