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First published online August 25, 2005
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2005-0148v1
24/2/368    most recent
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Submitted on April 1, 2005
Accepted on August 12, 2005

Original Article

Cell Size Correlates with Phenotype and Proliferative Capacity in Human Corneal Epithelial Cells

Cintia S. De Paiva 1, Stephen C. Pflugfelder 1, De-Quan Li 1*

1 Baylor College of Medicine, Houston, Texas

* To whom correspondence should be addressed. E-mail: dequanl{at}bcm.edu.


  Abstract

This study investigated whether cell size correlates with phenotype and proliferative capacity in human corneal epithelial cells. Primary cultured human corneal epithelial cells were sorted by flow cytometry based on Forward Scatter profile in comparison to the profile of beads of known size. Four fractions (A, B, C and D) of cells ranging in size from 10-16, 17-23, 24-30 and ≥31 µm in diameter, respectively, were collected to evaluate their BrdU-label retention properties, cell phenotype, and clonal growth capacity on a 3T3 fibroblast feeder layer. Among these 4 populations, cell size was shown to positively correlate to the expression of the differentiation markers, Keratin (K) 3, K12 and involucrin, and inversely to the levels of stem cell associated markers, {Delta}Np63 and ABCG2, and to colony forming efficiency (CFE) and growth capacity. The population A with the smallest size, accounting for 11.0 ± 4.5% of the entire population, contained the greatest number of BrdU label-retaining slow-cycling cells, displayed the highest percentage of cells immuno-positive to p63 and ABCG2 and negative to K3 and involucrin, expressed the highest levels of {Delta}Np63 and ABCG2 mRNA, and the lowest levels of K3, K12 and involucrin, and possessed the highest CFE and growth capacity. These findings suggest that the cell size correlates with cell differentiation phenotypes and proliferative capacity in human corneal epithelial cells. The smallest cells in population A appear to be enriched for putative stem cells, and small cell size may represent one of the important properties of adult corneal epithelial stem cells.

Key Words. cornea, limbus, epithelium, stem cells, cell size, BrdU




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