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K. Higa, S. Shimmura, N. Kato, H. Miyashita, T. Kawakita, Y. Ogawa, J. Shimazaki, K. Tsubota; N-Cadherin Function in Corneal Limbal Niche Model Using Feeder Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1774.
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© ARVO (1962-2015); The Authors (2016-present)
N-cadherin is a key component of the bone marrow stem cell niche. In a recent report, N-cadherin was reported to be expressed in the human limbal epithelial stem cell, and may be a critical cell-to-cell adhesion complex in the limbal epithelial niche. We reported that colony formation in 3T3 feeders might be used as a model of the limbal epithelial stem cell niche (2006ARVO). We demonstrated whether N-cadherin was indeed functionally required as a component of the limbal epithelial stem cell niche.
In order to show the importance of N-cad in colony formation, we used siRNA technology to downregulate N-cadheirn expression in 3T3 feeder cells (N-cadlow 3T3). Control plasmid was used as control. Epithelial sheet was also cultured using the N-cadlow 3T3 duplex feeder system required to reproduce the limbal phenotype of basal K15 expression in cultivated sheets.
When 3T3 cells transfected with N-cad siRNA were used as feeders, both CFE and colony sized were significantly reduced compared to control plasmid transfected cells. Cultivated sheets using this duplex feeder system with N-cadlow 3T3 cells lost the orientation of K15+ cells in the basal epithelial layer, while control plasmid transfected 3T3 cells were able to maintain basal K15+ epithelial cells.
N-cad plays an important role in the maintenance of K15+ corneal epithelial progenitor cells in a corneal limbal niche model using feeder cells.
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