Corneal epithelial stem cells are believed to reside in the basal layer of the limbal epithelium, but no definitive cell surface markers have been identified. For keratinocytes, stem/progenitor cells are known to be enriched by cell surface markers, integrin alpha(6) and CD71, as a minor subpopulation which shows high integrin alpha(6) and low CD71 expressions (alpha(bri)(6)/CD71(dim)). In the present study, we investigated the possibility that corneal epithelial stem cells can be enriched by integrin alpha(6) and CD71. The alpha(bri)(6)/CD71(dim) cells were separated by fluorescence-activated cell sorting, as a minor subpopulation of the limbal epithelial cells. They were enriched for relatively small cells, showing a higher clonogenic capacity and expression of stem cell markers, but a lower expression of differentiation markers, compared to other cell populations. The cells were localized immunohistochemically in the basal region of the limbal epithelium. These results indicate that the alpha(bri)(6)/CD71(dim) subpopulation enriched corneal epithelial stem cells. (c) 2007 Elsevier Inc. All rights reserved.