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Original Article |
1 Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
2 Department of Ophthalmology, Osaka University Medical School, Osaka, Japan
* To whom correspondence should be addressed. E-mail: tokano{at}abmes.twmu.ac.jp.
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Abstract |
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Corneal epithelial (CE) stem cells are believed to reside in the basal layer of the limbal epithelium, but remain poorly understood due to the lack of an accepted in vivo reconstitution assay, as well as definitive markers for epithelial stem cells. It has been reported that side population (SP) cells with the ability to efflux the DNA-binding dye Hoechst 33342 have stem cell-like properties and that the SP phenotype accurately represents a quiescent and immature stem cell population in the adult bone marrow. In the present study, we investigated whether SP cells isolated from the limbal epithelium have stem cell-like properties. SP cells, separated by fluorescence activated cell sorting (FACS), comprise approximately 0.4% of all limbal epithelial cells and have markedly higher expression of the stem cell markers ABCG2, Bmi-1, and nestin, but no expression of markers for differentiated CE cells compared to non-SP (NSP) cells. Cell cycle and telomerase activity analyses revealed that SP cells are growth arrested and reside in the quiescent state. Moreover, limbal epithelial SP cells did not demonstrate proliferative capabilities under typical in vitro epithelial cell culture conditions using 3T3 feeder layers. These findings present the possibility that quiescent limbal epithelial SP cells may represent an extremely immature stem cell population compared to currently defined epithelial stem or progenitor cells.
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