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Y Ban, NJ Fullwood, LJ Cooper, T Nakamura, M Tsuzuki, S Kinoshita; Quantitative Ultrastructural Comparison Of Human Corneal Epithelial Cells Cultivated On Amniotic Membrane With And Without Air-lifting . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1671.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:To evaluate the effectiveness of the air-lifting technique in culturing corneal limbal epithelial cells for use in ocular surface reconstruction, by comparing intercellular space and desmosome number in cultivated corneal epithelium using transmission electron microscopy, and by comparing cultivated epithelium morphology via scanning electron microscopy. Methods:Small pieces of corneal limbal epithelium from donor corneas were cultivated for 4 weeks on acellular amniotic membrane (AM) with inactivated 3T3 fibroblasts. For the first 3 weeks, all cultures were submerged in SHEM medium; they were then divided into 2 groups. The air-lift (+) group was cultured for the fourth week using the air-lifting technique, with the epithelium surface exposed to air by lowering the medium level. The air-lift (-) group was submerged in the medium for the entire 4 weeks. Cultured corneal epithelium morphology was examined via transmission and scanning electron microscopy. Results:In culture, both groups formed 4-5 layer-thick, well-stratified epithelium. Statistically, there was a highly significant difference in intercellular space size and desmosome number between the air-lift (+) and (-) groups. Under scanning electron microscopy, the air-lift (-) group epithelial cell junctions appeared to have small holes or gaps at the cell-cell interface. In the air-lift (+) group, by contrast, the epithelial cells were closely attached to each other and had distinct cell boundaries. Conclusion:The air-lifting technique tightens cell-cell attachment and creates the superficial epithelial cell barrier. This technique is effective in culturing corneal epithelial cells for use in ocular surface reconstruction.
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