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- Title
A Novel Isolation Technique of Progenitor Cells in Human Corneal Epithelium Using Non-Tissue Culture Dishes.
- Authors
Yokoo, Seiichi; Yamagami, Satoru; Shimada, Takashi; Usui, Tomohiko; Sato, Taka-Aki; Amano, Shiro; Araie, Makoto; Hamuro, Junji
- Abstract
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.
- Subjects
STEM cells; CORNEA; TISSUE culture; EPITHELIUM; CELL differentiation; SPECTRUM analysis
- Publication
Stem Cells, 2008, Vol 26, Issue 7, p1743
- ISSN
1066-5099
- Publication type
Article
- DOI
10.1634/stemcells.2007-0866