A Novel Isolation Technique of Progenitor Cells in Human Corneal Epithelium Using Non-Tissue Culture Dishes
Article first published online: 24 APR 2008
Copyright © 2008 AlphaMed Press
Volume 26, Issue 7, pages 1743–1748, July 2008
How to Cite
Yokoo, S., Yamagami, S., Shimada, T., Usui, T., Sato, T.-a., Amano, S., Araie, M. and Hamuro, J. (2008), A Novel Isolation Technique of Progenitor Cells in Human Corneal Epithelium Using Non-Tissue Culture Dishes. STEM CELLS, 26: 1743–1748. doi: 10.1634/stemcells.2007-0866
- Issue published online: 2 JAN 2009
- Article first published online: 24 APR 2008
- Manuscript Accepted: 14 APR 2008
- Manuscript Received: 29 OCT 2007
- Corneal epithelium;
- Epidermal growth factor;
The existence of adult stem cells or progenitor cells in the human corneal epithelium (i.e., self-renewing squamous cells) has long been suggested, but these cells have not yet been isolated. Here we describe a novel isolation technique using non-tissue culture dishes to enrich progenitor cells, which are able to reconstitute a three-dimensional human corneal epithelial equivalent from single cells in serum-, feeder-, and bovine pituitary extract-free medium. These cells showed original tissue-committed differentiation, a high proliferative capacity, and limited self-renewal. Laminin-5 was measured by mass spectrometric analysis. Pretreatment of cells with anti-laminin-5 antibody demonstrated that laminin-5 was important in allowing corneal epithelial progenitor cells to adhere to non-tissue culture dishes. Hydrophilic tubes (used for cell collection throughout this study) are essential for efficient isolation of adherent corneal epithelial progenitor cells expressing laminin-5. These findings indicate that our new technique using non-tissue culture dishes allows the isolation of progenitor cells from human corneal limbal epithelium and that laminin-5 has a critical role in the adhesion of these cells.
Disclosure of potential conflicts of interest is found at the end of this article.