June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
The different binding properties of cultured human corneal endothelial cell subpopulations to Descemet’s membrane components
Author Affiliations & Notes
  • Munetoyo Toda
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Morio Ueno
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Asako Hiraga
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Kazuko Asada
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Takahiro Nakamura
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Michio Hagiya
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Naoki Okumura
    Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Noriko Koizumi
    Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Junji Hamuro
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shigeru Kinoshita
    Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Footnotes
    Commercial Relationships Munetoyo Toda, None; Morio Ueno, Santen Pharmaceutical Co. (P), Senju Pharmaceutical Co. (P); Asako Hiraga, None; Kazuko Asada, None; Takahiro Nakamura, None; Michio Hagiya, JCR Pharmaceuticals Co. (E); Naoki Okumura, JCR Pharmaceuticals Co. (P), Senju Pharmaceutical Co. (P); Noriko Koizumi, Senju Pharmaceutical Co. (P), JCR Pharmaceuticals Co. (P); Junji Hamuro, None; Shigeru Kinoshita, Otsuka Pharmaceutical Co. (C), Santen Pharmaceutical Co. (P), Senju Pharmaceutical Co. (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1144. doi:
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      Munetoyo Toda, Morio Ueno, Asako Hiraga, Kazuko Asada, Takahiro Nakamura, Michio Hagiya, Naoki Okumura, Noriko Koizumi, Junji Hamuro, Shigeru Kinoshita; The different binding properties of cultured human corneal endothelial cell subpopulations to Descemet’s membrane components. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1144.

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

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Abstract

Purpose: Human corneal endothelial cells (HCECs) have poor proliferative ability under in vitro culture conditions. The tendency to enter into cell senescence or phase transition (epithelial-mesenchymal transition (EMT), cell senescence, and fibrosis) during cultivation produces different subpopulations from HCECs. To date, we have developed definitive subpopulations based on their cell surface markers. To clarify the adherent properties of these subpopulations, we compared the binding ability of cultured HCEC subpopulations to major Descemet’s membrane components that distribute to the endothelial face; i.e., laminin-511, -411, Type IV collagen, and proteoglycans.

Methods: Each subpopulation was prepared by controlling the culture conditions or by using magnetic cell separation, and then confirmed by staining with several cell surface markers. To examine the binding abilities of HCEC subpopulations, the cells were added to 96-well culture plates immobilized with collagens, laminins, or proteoglycans, and the plates were then centrifuged. The attached cells were then evaluated under a phase contrast microscope.

Results: Both the fully differentiated mature HCEC subpopulation and the EMT-phenotype subpopulation were found to be attached to laminin or collagen coated plates. Among the examined laminins, HCEC subpopulations were most strongly bound to laminin-511 and weakly bound to Perlecan, Agrin, and TSP-1. Interestingly, the binding properties to laminins were different among these subpopulations. Although the level of attached cells to the laminin-411 coated plate was the same among the HCEC subpopulations, the fully differentiated mature HCEC subpopulation was significantly more tightly bound to laminin-511 than was the EMT-phenotype subpopulation.

Conclusions: Our results suggest that the binding ability of HCECs to the major Descemet’s membrane components is distinct among subpopulations of cultivated HCECs. Moreover, the simple methods used in this study are effective for evaluating the interaction between HCECs and extracellular matrix components.

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