June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Cell Line of Fuchs' Corneal Dystrophy Produces an Abnormal Extracellular Matrix
Author Affiliations & Notes
  • Leona Ho
    Biomedical Engineering, Doshisha University, Kyoto, Japan
  • Naoki Okumura
    Biomedical Engineering, Doshisha University, Kyoto, Japan
  • EunDuck Kay
    Biomedical Engineering, Doshisha University, Kyoto, Japan
  • Kenta Yamasaki
    Biomedical Engineering, Doshisha University, Kyoto, Japan
  • Satoshi Kawasaki
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Theofilos Tourtas
    Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany
  • Ursula Schlotzer-Schrehardt
    Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany
  • Friedrich Kruse
    Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany
  • Shigeru Kinoshita
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Noriko Koizumi
    Biomedical Engineering, Doshisha University, Kyoto, Japan
  • Footnotes
    Commercial Relationships Leona Ho, None; Naoki Okumura, None; EunDuck Kay, None; Kenta Yamasaki, None; Satoshi Kawasaki, None; Theofilos Tourtas, None; Ursula Schlotzer-Schrehardt, None; Friedrich Kruse, None; Shigeru Kinoshita, Senju Pharmaceutical Co (P), Santen Pharmaceutical Co (P), Otsuka Pharmaceutical Co (C), Alcon (R), AMO (R), HOYA (R); Noriko Koizumi, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1680. doi:
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      Leona Ho, Naoki Okumura, EunDuck Kay, Kenta Yamasaki, Satoshi Kawasaki, Theofilos Tourtas, Ursula Schlotzer-Schrehardt, Friedrich Kruse, Shigeru Kinoshita, Noriko Koizumi; Cell Line of Fuchs' Corneal Dystrophy Produces an Abnormal Extracellular Matrix. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1680.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Abnormal deposition of extracellular matrix (ECM), such as corneal guttae, causes visual disturbance in patients with Fuchs’ endothelial corneal dystrophy (FECD), a chronic corneal disease that is known to cause blindness. To date, no pharmaceutical intervention is available for the treatment of patients afflicted with FEDC. The purpose of this study was to establish a cellular in vitro model that can be used for the potential development of a therapy to treat patients with FECD.

Methods: Human corneal endothelial cells (HCECs) were cultured from endothelial-Descemet membrane lamellae obtained from 3 FECD patients during Descemet's membrane endothelial keratoplasty (DMEK) after obtaining informed written consent from each patient. HCECs cultured from 3 individual normal donor corneas were used as a control. The FECD-derived HCECs and the control HCECs were then immortalized by SV40 and hTERT to produce iFECD and iHCEC cell lines, respectively. The gene expression levels of ECM components were then analyzed by TaqMan® real-time PCR. To elucidate ECM production, iFECD and iHCEC cells were cultured in a Transwell® culture system, and ECM deposition was then analyzed by hematoxylin-eosin (HE) staining and immunohistochemistry after 2 weeks.

Results: Real-time PCR revealed a significantly increased production of type I collagen, type IV collagen, and fibronectin (p<0.05) in iFECD cells in comparison to iHCEC cells. In addition, the gene expression of ZEB2, an epithelial to mesenchymal transition transcription factor, was also found to be significantly increased (p<0.01) in iFEDC in comparison to iHCEC cell lines. HE staining showed that a significant thicker ECM was produced by iFECD cells than by iHCEC cells (6.65±0.82μm and 3.14±0.64μm, respectively) (p<0.01). Immunostaining also showed a greater intensity of staining for ECM proteins, such as collagen and fibronectin, in iFECD cells.

Conclusions: We established a cellular model of FECD associated with an abnormal production of ECM. These findings may prove valuable as a research tool that can be used for the potential development of a novel treatment option for patients with FECD.

Keywords: 481 cornea: endothelium • 480 cornea: basic science  
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