April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
N-Cadherin Function in Corneal Limbal Niche Model Using Feeder Cells
Author Affiliations & Notes
  • K. Higa
    Ophthalmology, Tokyo Dental College, Ichikawa-shi, Japan
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • S. Shimmura
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • N. Kato
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • H. Miyashita
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • T. Kawakita
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • Y. Ogawa
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • J. Shimazaki
    Ophthalmology, Tokyo Dental College, Ichikawa-shi, Japan
  • K. Tsubota
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • Footnotes
    Commercial Relationships  K. Higa, None; S. Shimmura, None; N. Kato, None; H. Miyashita, None; T. Kawakita, None; Y. Ogawa, None; J. Shimazaki, None; K. Tsubota, None.
  • Footnotes
    Support  Advanced and Innovational Research program in Life Sciences from the Ministry of Education
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1774. doi:
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      K. Higa, S. Shimmura, N. Kato, H. Miyashita, T. Kawakita, Y. Ogawa, J. Shimazaki, K. Tsubota; N-Cadherin Function in Corneal Limbal Niche Model Using Feeder Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1774.

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

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Abstract

Purpose: : N-cadherin is a key component of the bone marrow stem cell niche. In a recent report, N-cadherin was reported to be expressed in the human limbal epithelial stem cell, and may be a critical cell-to-cell adhesion complex in the limbal epithelial niche. We reported that colony formation in 3T3 feeders might be used as a model of the limbal epithelial stem cell niche (2006ARVO). We demonstrated whether N-cadherin was indeed functionally required as a component of the limbal epithelial stem cell niche.

Methods: : In order to show the importance of N-cad in colony formation, we used siRNA technology to downregulate N-cadheirn expression in 3T3 feeder cells (N-cadlow 3T3). Control plasmid was used as control. Epithelial sheet was also cultured using the N-cadlow 3T3 duplex feeder system required to reproduce the limbal phenotype of basal K15 expression in cultivated sheets.

Results: : When 3T3 cells transfected with N-cad siRNA were used as feeders, both CFE and colony sized were significantly reduced compared to control plasmid transfected cells. Cultivated sheets using this duplex feeder system with N-cadlow 3T3 cells lost the orientation of K15+ cells in the basal epithelial layer, while control plasmid transfected 3T3 cells were able to maintain basal K15+ epithelial cells.

Conclusions: : N-cad plays an important role in the maintenance of K15+ corneal epithelial progenitor cells in a corneal limbal niche model using feeder cells.

Keywords: cornea: endothelium • cell adhesions/cell junctions • RNAi 
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