IOVS Journal of Neurophysiology
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(Investigative Ophthalmology and Visual Science. 2007;48:4519-4526.)
© 2007 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-0567
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Cultivated Corneal Endothelial Cell Sheet Transplantation in a Primate Model

Noriko Koizumi,1,2 Yuji Sakamoto,3 Naoki Okumura,2 Norio Okahara,4 Hideaki Tsuchiya,4 Ryuzo Torii,4 Leanne J. Cooper,2 Yuriko Ban,2 Hidetoshi Tanioka,2 and Shigeru Kinoshita2

1From the Research Center for Regenerative Medicine, Doshisha University, Kyoto, Japan; the 2Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan; the 3Research Laboratory, Senju Pharmaceutical Co., Ltd., Kobe, Japan; and the 4Research Center for Animal Life Science, Shiga University of Medical Science, Ohtsu, Japan.

PURPOSE. To examine the feasibility of cultivated corneal endothelial cell transplantation in a primate model.

METHODS. Monkey corneal endothelial cells (MCECs) obtained from three cynomolgus monkeys were cultivated, with subcultures grown on collagen type I carriers for 4 weeks. The corneal endothelium of the right eye of six monkeys was mechanically scraped, after which a cultivated MCEC sheet was brought into the anterior chamber of four eyes and fixed to Descemet’s membrane by air. As the control, a collagen sheet without MCECs was transplanted into one eye of one monkey, and a suspension of cultivated MCECs was injected into the anterior chamber in one eye.

RESULTS. Cultivated MCECs produced a confluent monolayer of closely attached hexagonal cells that showed both ZO-1 and Na+-K+ ATPase expression. In the early postoperative period MCEC sheets were attached to Descemet’s membrane and corneal clarity was recovered. The recovered clarity was accompanied by a decrease in corneal thickness. Fluorescein DiI labeled donor corneal endothelial cells were detected on the host cornea on postoperative day 7. Six months after transplantation MCEC-transplanted corneas remained clear, and hexagonal cells were observed by in vivo specular microscopy with a density of 1992 to 2475 cells/mm2. Control eyes showed irreversible bullous keratopathy that precluded pachymetry and specular microscopy.

CONCLUSIONS. A model of cultivated corneal endothelial transplantation for corneal endothelial dysfunction was established in primates whose corneal endothelial cells have less proliferative capacity in vivo. Our results suggest that this is a useful model for long-term observation in advance of the future clinical application of cultivated corneal endothelial transplantation.






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Copyright © 2007 by the Association for Research in Vision and Ophthalmology