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G.B. Melo, M.A. Glória, E.F. Haapalainen, J.A. P. Gomes; Comparison Between Amniotic Membrane With and Without Epithelium as a Support for Human Limbal Epithelial Cells Cultured ex–vivo . Invest. Ophthalmol. Vis. Sci. 2006;47(13):3027.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the efficacy and ultrastructural aspects of human limbal epithelial cells cultured on amniotic membrane (AM) with and without epithelium.
Limbal epithelial cell cultures were established from cadaveric corneo–scleral rim explants derived from 6 different donors. The explants from each donor were placed in 35mm cell culture dishes under 3 different conditions: over the bottom of one wheel (control), on human preserved amniotic membrane with epithelium and on human preserved AM deepithelialized with trypsin (2 mg/ml at 1:250). The epithelial cell migration was evaluated by two observers under phase contrast microscopy. After 15 days, the AM with cells cultures were removed and submitted to scanning and transmission electron microscopy to check for epithelial migration and adhesion.
All cells cultures from the controls grew until reaching confluence. All but one denuded AM presented epithelial migration. The growing of epithelial cell over AM with epithelium was observed only in 1 group.
Using this model, denuded AM with the use of trypsin appeared to be the best support for epithelial cell migration and adhesion comparing to the AM with epithelium. Removal of AM epithelial seems to be an important step for establishig limbal epithelial cells cultured on AM.
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