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S.C. Yiu, D. Wasilewski, D. Stevenson, E. Barron, A.R. Rodriguez, D.T. Woodley, M.D. Trousdale, J.C. Song, J.A. Irvine, R.E. Smith; High Expression Level of ß-1 Integrins in Expansion of Human Corneolimbal Explants . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1351.
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Purpose: A previous report from our laboratory (Yiu et al, Invest Ophthalmol Vis Sci; 2001, 5477, vol. 42(4)) showed high ß-1-integrin expression in the human corneolimbal stem cell compartment. This suggests that ß-1 integrin is a potential marker for corneal stem cells. If this unique property persists in primary culture, it may be possible to isolate these cells individually and expand them to confluence to obtain a pure population of stem cells. The present study is to determine whether cultured corneolimbal epithelial cells continue to express high levels of ß1 integrin. Methods: Human corneal donor rims were obtained from corneal transplant surgery. Explants, each 2 mm x 2 mm, were obtained from the corneolimbal area and placed in a sterile petri dish with growth media (EpiLife, Cascade Biologic). Cultured epithelial cells were grown to confluence and fixed with 4% paraformaldehyde. The cells were then stained with ß1 and α6 and with other antibodies. Staining profiles were detected and analyzed by confocal microscopy. Results:Clusters of cells was stained brightly with ß-1 integrins; others were stained brightly with α-6 integrins. But not all cells stained brightly with either ß-1 integrins or α-6 integrins. This differential expression of cell surface proteins supports the findings of our previous whole tissue study. Conclusions: Our results demonstrate that the primary culture of corneolimbal explants retained characteristics similar to cells located in the human corneolimbal stem cell compartment. The bright ß-1-integrin staining of clusters of cells suggested that the cells could be putative corneal stem cells; the bright α-6-integrin staining of the other clusters suggested that they could be transient amplifying cells. This differential staining pattern will enable us to isolate these cells individually and further delineate their characteristics.
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