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L. Germain, P. Carrier, A. Deschambeault, M. Talbot, S.L. Guerin, F.A. Auger; FIBROBLASTS MODULATE DIFFERENTIATION AND STRATIFICATION OF EPITHELIAL CELLS ON RECONSTRUCTED HUMAN CORNEA THROUGH THE PRODUCTION OF SOLUBLE FACTORS . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1451.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Interactions between stromal cells and epithelial cells are present during processes such as development and maintenance of cornea and skin. To investigate the role of stromal cells on the differentiation and stratification of the epithelium, a recently developed human self–assembled corneal model produced by tissue engineering was used. To better evaluate whether the factors involved in this interaction were soluble, the effect of conditioned media was evaluated. Methods: We have reconstructed four types of tissues (according to the following combinations): corneal fibroblasts/corneal epithelial cells, corneal fibroblasts/skin epithelial cells, skin fibroblasts/skin epithelial cells, skin fibroblasts/corneal epithelial cells. Fibroblasts isolated from skin or cornea were cultured in the presence of ascorbic acid to produce sheets on which epithelial cells were then seeded and cultured. Then, a conditioned media was prepared from corneal and skin fibroblast’s monolayer cultures and added to the media in which reconstructed tissues are cultured. Histological studies were performed. Results: A higher differentiation status was observed in the reconstructed epidermis (15 to 20 cell layers including a stratum corneum) compared to the reconstructed cornea (6 cell layers, no stratum corneum), as observed in situ. The other two reconstructed tissues gave intermediate differentiation levels with thinner or absent stratum corneum. The media conditioned by a subconfluent monolayer of corneal fibroblasts was able to decrease the number of cell layers in the epithelium of the reconstructed skin and also in the skin fibroblasts/corneal epithelial cells reconstructed tissue. In contrast, the media conditioned by skin fibroblasts was able to increase the differentiation status of the reconstructed cornea and corneal fibroblasts/skin epithelial cells reconstructed tissue. Work is in progress to identify the factors possibly involved in the regulation of epithelium thickness. Conclusions: These results demonstrate that soluble factors secreted by skin fibroblasts differs from those produced by corneal fibroblasts. Moreover, these soluble factors produced by mesenchymal cells modulate the stratification and differentiation of epithelial cells on the reconstructed tissues.
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