Pluripotent Stem Cell-Derived Corneal Endothelial Cells Form a Functional Corneal Endothelium in Canines.

PURPOSE: We previously reported that cryopreserved human embryonic stem cell (hESC)-derived corneal endothelial cells (CECs) can form a functional corneal endothelium (CE) on denuded Descemet's membrane (DM) in rabbits and monkeys. Here, we extend the utility of cryopreserved CECs to develop a possible treatment for canines with corneal endothelial dysfunction.

METHODS

We developed a corneal endothelial dysfunction canine model (n = 2) by mechanical scraping of central (8 mm diameter) CE. To regenerate the CE, the canines received an injection of cryopreserved hESC-derived CECs in the anterior chamber of the eye, followed by being placed in a cornea-down position to allow the injected cells to settle on denuded DM. The structural integrity, pachymetry, CE cell density and morphology of the regenerated and the CE of the untreated eyes were examined by optical coherence tomography (OCT), and a specular microscope. Immunohistochemical (IHC) analysis of the regenerated and resident CE was completed with a human-specific nucleoli antibody and other antibodies.

RESULTS

The corneas of the cryopreserved hESC-derived CEC-injected eyes developed corneal edema within 24 hours after scraping of the CE and cell injection but regained transparency in one to two weeks after the CEC injection. OCT revealed anatomically normal corneas with pachymetry of the CEC-injected eyes similar to the untreated left eyes, and specular microscopy examination illustrated regenerated CE exhibiting hexagonal/polygonal cells with CEC density of the regenerated CE comparable to the CE of the untreated eye. IHC analysis confirmed the regeneration of a functional CE resulting from the settlement of injected cryopreserved hESC-derived CECs on the denuded DM.

CONCLUSIONS

The results confirm the regeneration of a functional CE in canines resulting from the settlement of cryopreserved hESC-derived CECs on the denuded DM.

TRANSLATIONAL RELEVANCE: Our data confirm the efficacy of cryopreserved pluripotent stem cell-derived CECs as a therapeutic option to regenerate a functional CE in canines with corneal endothelial dysfunction.