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Y Ishino, Y Sano, T Nakamura, K Endo, M Tsuzuki, N Tanifuji, S Kinoshita; Amniotic Membrane as Carrier for Cultured Corneal Endothelium in vitro . Invest. Ophthalmol. Vis. Sci. 2002;43(13):854.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:Previous studies have shown that human corneal endothelial cells (HCEC) can be expanded in vitro, which indicates that cultured HCEC can be transplanted in endothelial diseases, such bullous keratopathy or endothelial degeneration, without damaging stroma or epithelium. To transplant HCEC in vivo, a substrate that carries HCEC is required. Since amnitotic membrane (AM) is reportedly a good carrier for cultivated corneal epithelium, we wished to examine the feasibility of using amniotic membrane as a carrier forHCEC. Methods:Healthy corneal tissues were obtained from an eye bank in United States. Descemet`s membranes were dissected from the stroma, and endothelial cell suspensions were obtained by dispase treatment. Endothelial cells were seeded in wells of a 24-well plate coated with collagen IV for primary culture. After 3 (P3) or 6 (P6) culture passages, 1.2x105/ml (P6 HCEC) or 4.5x104;/ml (P3 HCEC) of cells were seeded on AM denuded of epithelial cells or on collagen IV-coated wells of a 12-well plate. At 2 weeks, endothelial cell densities were measured and their averages calculated. Results:On a collagen IV-coated dish, cell densities at 2 weeks of culture were 805±122/mm2 (P6 HCEC) and 777±86/mm2 (P3, HCEC). On AM, the figures were 867±245/mm2 (P6, HCEC) and 646±189/mm2 (P3, HCEC). There were no significant differences in cell densitiy between cultures on collagen IV and AM. Conclusion:Our data show that HCEC densities on AM are equivalent to those on collagen IV-coated wells. Though higher cell densities are required for in vivo transplantation, these results indicate that AM may be a candidate carrier of cultivated HCEC for HCEC transplantation.
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