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Noriko Koizumi, Nigel J. Fullwood, George Bairaktaris, Tsutomu Inatomi, Shigeru Kinoshita, Andrew J. Quantock; Cultivation of Corneal Epithelial Cells on Intact and Denuded Human Amniotic Membrane. Invest. Ophthalmol. Vis. Sci. 2000;41(9):2506-2513.
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purpose. Surgery to reconstruct the ocular surface is greatly facilitated by the
use of amniotic membrane, either as a biologic drape or, more recently,
as a substrate for the transplantation of cultivated corneal epithelial
cells. This study was designed to compare the usefulness of intact and
denuded human amniotic membranes as a substrate for corneal epithelial
methods. Small (3-mm-diameter) biopsy specimens of superficial cornea including
epithelium were excised from the central and limbal regions in rabbits.
They were cultured on human amniotic membrane with or without amniotic
epithelial cells and examined by light, scanning electron, and
transmission electron microscopy.
results. Cellular outgrowth from the central explants (n = 10)
after 14 days in culture measured 1.82 ± 2.62
mm2 on intact amniotic membrane and 131.83 ± 28.31 mm2 on denuded amniotic membrane. In
contrast, outgrowths from the limbal explants (n = 10)
at the same time measured 4.58 ± 4.56 and 505.39 ± 134.20
mm2 on intact and denuded amniotic membranes,
respectively. The leading edges of the outgrowths on intact amniotic
membrane were much less uniform than those on denuded amniotic
membrane, and, in the former, corneal epithelial cells appeared to
migrate over the top of amniotic epithelial cells. Limbal cells
cultivated on denuded amniotic membrane formed a nicely stratified
layer that adhered well to the underlying amniotic membrane.
conclusions. Denuded amniotic membrane appears to be an excellent substrate for the
cultivation of corneal epithelial cells, with a view to
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