Corneal Epithelial Cultures Generated from Organ-Cultured Limbal Tissue: Factors Influencing Epithelial Cell Growth.

Autor: Zito-Abbad, Eleonora, Borderie, Vincent M., Baudrimont, Marie, Bourcier, Tristan, Laroche, Laurent, Chapel, Catherine, Uzel, Jean-Louis
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Zdroj: Current Eye Research; May2006, Vol. 31 Issue 5, p391-399, 9p, 2 Black and White Photographs, 3 Graphs
Abstrakt: Purpose : To explore the in vitro proliferative potential of human limbal epithelial cells after 31°C organ-culture storage and to investigate putative factors influencing it. Methods : 185 cultures of limbal explants were carried-out either from full-thickness explants (n = 102) or from enzymatically dissociated cells (n = 83) seeded on a feeder layer of human keratocytes. Epithelial outgrowth was assessed by phase contrast microscopy using a computerized image analysis software. Cell phenotype was evaluated by transmission electron microscopy and immunocytology. Univariate and multivariate analysis were performed to determine factors influencing epithelial growth in culture. Results : An epithelial outgrowth of 100 square mm or more was observed in 52% of cultures, (average growth area: 440 +/- 256 mm at three weeks). Corneal epithelial phenotype was confirmed by transmission electron microscopy, and cytokeratin pattern. Cytokeratine 19, deltaNp63, nestin and vimentin positive staining revealed undifferenciated epithelial cells in both explant and cell suspension cultures at three weeks. Short death to cornea retrieval time (p [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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