March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Cell-injection Therapy using Cultivated Human Corneal Endothelial Cells in a Primate Model
Author Affiliations & Notes
  • Noriko Koizumi
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural Univ of Med, Kyoto, Japan
  • Naoki Okumura
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural Univ of Med, Kyoto, Japan
  • Yuji Sakamoto
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Kenta Yamasaki
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Makiko Nakahara
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Ryuzo Torii
    Research Center for Animal Life Science, Shiga University of Medical Science, Otsu, Japan
  • Morio Ueno
    Ophthalmology, Kyoto Prefectural Univ of Med, Kyoto, Japan
  • Junji Hamuro
    Ophthalmology, Kyoto Prefectural Univ of Med, Kyoto, Japan
  • Shigeru Kinoshita
    Ophthalmology, Kyoto Prefectural Univ of Med, Kyoto, Japan
  • Footnotes
    Commercial Relationships  Noriko Koizumi, None; Naoki Okumura, None; Yuji Sakamoto, None; Kenta Yamasaki, None; Makiko Nakahara, None; Ryuzo Torii, None; Morio Ueno, None; Junji Hamuro, None; Shigeru Kinoshita, None
  • Footnotes
    Support  the Funding Program for Next Generation World-Leading Researchers from the Cabinet Office in Japan (Koizumi LS117), MEXT, Highway Program for Realization of Regenerative Medicine (Kinoshita)
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 1738. doi:
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      Noriko Koizumi, Naoki Okumura, Yuji Sakamoto, Kenta Yamasaki, Makiko Nakahara, Ryuzo Torii, Morio Ueno, Junji Hamuro, Shigeru Kinoshita; Cell-injection Therapy using Cultivated Human Corneal Endothelial Cells in a Primate Model. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1738.

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

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Abstract

Purpose: : To investigate the feasibility of corneal endothelial reconstruction by a cell-injection therapy using cultivated human corneal endothelial cells (HCECs) in a non-human primate model.

Methods: : Monkey corneal endothelium was intensively scraped off up to the peripheral area to make a corneal endothelial dysfunction model. A 2.0 x 105 amount of cultivated HCECs (passage 3) suspended in OptiMEM-I supplemented with 100μM of selective rho kinase (ROCK)-inhibitor Y-27632 was injected into the anterior chamber of 2 eyes of 2 animals (Y-27632(+) group). The same procedure was performed without using ROCK-inhibitor Y-27632 in 2 eyes of 2 animals (Y-27632(-) group). The eyes of each animal were then kept in the face-down position for 3 hours. In the control group, endothelial cells were scraped and HCECs were not injected (2 eyes). Slit-lamp examinations and corneal thickness- and intraocular pressure measurements were then performed for up to 2.5 months, followed by immunohistochemical analysis.

Results: : Two eyes in the control group demonstrated severe corneal edema and did not recover corneal clarity. In contrast, the 4 eyes that received cell-injection therapy showed improved corneal clarity. Corneal clarity recovered faster and the corneal thickness at 2.5 months after treatment was thinner in the Y-27632(+) eyes compared to the Y-27632(-) eyes (507μm and 961μm, respectively). A homogeneous monolayer of polygonal cells expressing ZO-1 and Na+/K+-ATPase was reconstructed in the Y-27632(+) group, and the corneal endothelial cell density was much higher in the Y-27632(+) group compared to the Y-27632(-) group. None of the eyes showed intraocular pressure elevation or an immunological rejection.

Conclusions: : The findings of this present study indicate that cell-injection therapy using ROCK-inhibitor Y-27632 might be a clinically applicable procedure for corneal endothelial dysfunction.

Keywords: cornea: endothelium • cornea: basic science • transplantation 
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