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International Journal of Ophthalmic PathologyISSN: 2324-8599

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Research Article, Int J Ophthalmic Pathol Vol: 1 Issue: 2

Cultured Human Keratocytes from the Limbus and Cornea Both Express Epithelial Cytokeratin 3: Possible Mesenchymal-Epithelial Transition

Giuseppina Perrella1, Cathryn Anne Scott2*, Renza Spelat1, Paolo Brusini3, Federica D’Aurizio2, Ilaria De Pol1 and Harminder Singh Dua4
1Institute of General Pathology, Department of Medical and Biological Sciences, Faculty of Medicine, University of Udine, Italy
2Institute of Anatomic Pathology, Department of Medical and Biological Sciences, Faculty of Medicine, University of Udine, Italy
3Department of Ophthalmology, Azienda Ospedaliero-Universitaria S. Maria della Misericordia, Udine, Italy
4Department of Ophthalmology and Visual Sciences, University of Nottingham, Nottingham, United Kingdom
Corresponding author : Dr. Cathryn Anne Scott
Institute of Anatomic Pathology, Department of Medical and Biological Sciences, Faculty of Medicine, University of Udine, c/o Azienda Ospedaliero-Universitaria S. Maria della Misericordia, via Pieri, 33100 Udine, Italy
Tel: +390432559404; Fax: +390432559420
E-mail: [email protected]
Received: July 28, 2012 Accepted: September 17, 2012 Published: September 20, 2012
Citation: Perrella G, Scott CA, Spelat R, Brusini P, D’Aurizio F, et al. (2012) Cultured Human Keratocytes from the Limbus and Cornea Both Express Epithelial Cytokeratin 3: Possible Mesenchymal-Epithelial Transition. Int J Ophthalmic Pathol 1:2. doi:10.4172/2324-8599.1000101

Abstract

The corneal limbus is the repository of epithelial stem cells (SC) that sustain the turnover of corneal epithelial cells. The limbus stroma contains mesenchymal SC that generates stromal keratocytes. Mesenchymal-epithelial transition is a phenomenon wherein cells of mesenchymal phenotype can transdifferentiate to epithelial phenotype. Our aim was to study whether limbal keratocytes, cytokeratin 3 (CK3) negative, could be induced to transdifferentiate into CK3 positive cells.

Human keratocytes were isolated from the limbus and cornea of cadaver donors, cultured and evaluated for CD34, CK3 and vimentin expression by immunofluorescence and RTPCR and for keratocan by RT-PCR.

All cells regardless of site expressed vimentin and some also expressed CD34 and CK3. Double immunofluorescence revealed three subpopulations: CK3−/CD34+, CK3+/CD34+ and CK3+/CD34−. Total CD34 cell yield was higher in the limbus with a peak time to confluence (TTC) of more than 30 days. Total CK3 cell yield was greater in the cornea with a peak TTC of less than 30 days. Increasing donor age corresponded to a decreased CD34 yield and an increased CK3 yield. CK3−/CD34+ and CK3+/CD34− cells behaved similarly to total CD34 and CK3 cells in relation to age, site and TTC while CK3+/CD34+ cells showed intermediate features. Keratocan was present in corneal samples.

Keywords: Limbus; Cornea; Keratocyte; Cytokeratin 3; CD34; Keratocan

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