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A. R. Harris, C. Mason, J. T. Daniels; Intermediate Oxygen Tensions Promote Maintenance and Expansion of the Limbal Epithelial Stem Cell Phenotype. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1781.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effect of low oxygen on the maintenance of the limbal epithelial stem cell phenotype.
Limbal epithelial cells were isolated from rabbit corneas and cultured at various oxygen tensions between 21% and 2% in a co-culture system with Mitomycin C growth arrested 3T3 fibroblasts. The colony forming efficiency and proliferative ability of cells cultured in these different oxygen tensions were determined, and immunohistochemistry was used to detect expression of putative limbal epithelial stem cell markers such as ABCG2 and beta 1 integrin brightness.
An oxygen tension of 14% was optimal for the expansion of limbal epithelial stem cells in vitro, as indicated by higher colony forming efficiency, faster proliferation, smaller cell size, and the expression of putative limbal stem cell markers including ABCG2.
This study suggests that intermediate oxygen tensions promote the maintenance and expansion of rabbit limbal epithelial stem cells in a feeder co-culture system. Sub-atmospheric oxygen tensions are therefore recommended for the culture of rabbit limbal epithelial cells and may have relevance for the culture of human limbal epithelial cells for therapeutic application.
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