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Exp Eye Res. 2011 Jan;92(1):10-9. doi: 10.1016/j.exer.2010.10.004. Epub 2010 Oct 21.

Effects of fibroblast origin and phenotype on the proliferative potential of limbal epithelial progenitor cells.

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  • 1Discipline of Medical Sciences and Vision Improvement Domain, Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, Brisbane, QLD 4059, Australia. sl.ainscough@qut.edu.au

Abstract

The current study investigates potential differences in fibroblast phenotype across the anterior segment of the human eye with the aim to understanding factors that support the regenerative function of human limbal epithelial progenitor cells (LEPs) during wound healing. Separate cultures of fibroblasts were established from the cornea, limbus and sclera by growth in serum-supplemented medium. The resulting cultures were examined for potential differences in morphology and growth rate, as well as expression of CD34, CD45, CD90, CD141, CD271, vimentin and α-smooth muscle actin (α-sma). Finally, cultures were examined for their ability to support the growth of LEPs. While all cultures grew at a similar rate, scleral cultures often contained larger and more irregularly shaped cells which stained positive for α-sma. Western blotting confirmed a gradient of α-sma expression with lowest levels in corneal cultures. All three cultures stained positively for CD90 and vimentin, and were negative for CD34, CD45, CD141 and CD271. Only limbal or corneal irradiated fibroblasts supported the establishment of LEP cultures. While LEP colony forming efficiency and prominent expression of ABCG2, C/EBPδ and p63 was similar with either limbal or corneal fibroblasts, limbal fibroblasts supported significantly better growth. These results indicate that scleral fibroblasts have an increased capacity for myofibroblast formation which appears to negatively impact on their ability to support LEP growth. Superior growth of LEPs in the presence of limbal fibroblasts indicates a role for limbal fibroblasts in promoting the proliferation of limbal epithelium during wound healing.

PMID:
20970420
DOI:
10.1016/j.exer.2010.10.004
[PubMed - indexed for MEDLINE]
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