Here, we evaluate the efficacy of cryopreserved human embryonic stem cell (hESC)-derived corneal endothelial cells (CECs) to form a functional monolayer of corneal endothelium (CE) in rabbits and monkeys. We injected cryopreserved hESC-derived CECs into the anterior chamber of rabbits and monkeys either immediately after mechanical scraping of the central CE or a few days later when corneal edema developed. All preclinical models developed deturgesced and clear corneas following the injection of cryopreserved hESC-derived CECs and remained comparable to the corneas of the untreated eye. Confocal scanning microscopy confirmed an intact structure of hexagonal/polygonal cells and immunohistochemical analysis illustrated a monolayer expressing barrier and pump function proteins in the regenerated CE. The necropsy examination confirmed no remarkable change in multiple tissues assessed for teratoma formation. In conclusion, our data demonstrate the efficacy of cryopreserved hESC-derived CECs to form a functional CE on the denuded Descemet's membrane.

在这里，我们评估了冷冻保存的人胚胎干细胞 (hESC) 衍生的角膜内皮细胞 (CEC) 在兔和猴子体内形成功能性单层角膜内皮 (CE) 的功效。我们将冷冻保存的 hESC 衍生的 CEC 注射到兔子和猴子的前房中，或者在中央 CE 机械刮擦后立即或几天后角膜水肿发生时注射。在注射冷冻保存的 hESC 衍生的 CEC 后，所有临床前模型都形成了去角质和透明的角膜，并且与未经治疗的眼睛的角膜保持可比性。共聚焦扫描显微镜证实了六边形/多边形细胞的完整结构，免疫组织化学分析显示了在再生 CE 中表达屏障和泵功能蛋白的单层。尸检证实在评估畸胎瘤形成的多个组织中没有显着变化。总之，我们的数据证明了冷冻保存的 hESC 衍生的 CEC 在裸露的后弹力膜上形成功能性 CE 的功效。