Cultivation of corneal epithelial cells on intact and denuded human amniotic membrane

PURPOSE. Surgery to reconstruct the ocular surface is greatly facilitated b y the use of amniotic membrane, either as a biologic drape or, more recentl y, as a substrate for the transplantation of cultivated corneal epithelial cells. This study was designed to compare the usefulness of intact and denu ded human amniotic membranes as a substrate for corneal epithelial cell cul ture. METHODS. Small (3-mm-diameter) biopsy specimens of superficial cornea inclu ding epithelium were excised from the central and limbal regions in rabbits . They were cultured on human amniotic membrane with or without anmiotic ep ithelial cells and examined by light, scanning electron, and transmission e lectron microscopy. RESULTS. Cellular outgrowth from the central explants (n = 10) after 14 day s in culture measured 1.82 +/- 2.62 mm(2) on intact amniotic membrane and 1 31.83 +/- 28.31 mm(2) 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 mm(2) on intact and denuded amniotic membranes, respe ctively. The leading edges of the outgrowths on intact amniotic membrane we re much less uniform than those on denuded amniotic membrane, and, in the f ormer, corneal epithelial cells appeared to migrate over the top of amnioti c epithelial cells. Limbal cells cultivated on denuded amniotic membrane fo rmed 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 transplant ation.