Cultivated corneal endothelial cell sheet transplantation in a primate model.
OBJECTIVE
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/mm(2). 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.
Koizumi
Noriko
N
Research Center for Regenerative Medicine, Doshisha University, Kyoto, Japan. nkoizumi@ophth.kpu-m.ac.jp
Sakamoto
Yuji
Y
Okumura
Naoki
N
Okahara
Norio
N
Tsuchiya
Hideaki
H
Torii
Ryuzo
R
Cooper
Leanne J
LJ
Ban
Yuriko
Y
Tanioka
Hidetoshi
H
Kinoshita
Shigeru
S
D000818:Animals; D000867:Anterior Chamber / Q000601:surgery*; D002448:Cell Adhesion; D002452:Cell Count; D017690:Cell Transplantation; D002478:Cells, Cultured; D003886:Descemet Membrane / Q000601:surgery*; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006085:Graft Survival / Q000502:physiology; D008252:Macaca fascicularis; D008565:Membrane Proteins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D023421:Models, Animal; D010750:Phosphoproteins / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014184:Transplantation, Homologous; D062826:Zonula Occludens-1 Protein
D005456:Fluorescent Dyes; D008565:Membrane Proteins; D010750:Phosphoproteins; D062826:Zonula Occludens-1 Protein; D000254:Sodium-Potassium-Exchanging ATPase