人类多能干细胞（hPSC）的研究显示出基于细胞的再生医学取得了巨大进展。角膜内皮功能障碍与角膜内皮细胞（CEC）的丢失和退化有关，因此细胞替代成为一种有前途的治疗策略。然而，针对这种细胞疗法对 hPSC 衍生的 CEC 进行全面的临床前评估仍然是一个挑战。在这里，我们定义了一种适应的分化方案，以一致、有效地从临床级人胚胎干细胞 (hESC) 与异种细胞生成诱导角膜内皮细胞 (iCEC) -不含培养基和制造的冷冻保存的 iCEC。细胞表达高水平的典型 CEC 标记物，并在体外表现出 iCEC 典型的跨内皮电位特性。经过严格的质量控制措施后，符合所有释放标准的细胞可用于体内研究。我们发现免疫缺陷小鼠的移植物没有过度生长或致瘤性。移植到兔子模型后，存活的 iCEC 改善了水肿并恢复了角膜混浊。我们的工作为生成 iCEC 提供了一种有效的方法，并证明了 iCEC 在疾病建模中的安全性和有效性。因此，临床级 iCEC 是未来角膜内皮功能障碍临床治疗的可靠来源。© 2023。作者。

Research on human pluripotent stem cells (hPSCs) has shown tremendous progress in cell-based regenerative medicine. Corneal endothelial dysfunction is associated with the loss and degeneration of corneal endothelial cells (CECs), rendering cell replacement a promising therapeutic strategy. However, comprehensive preclinical assessments of hPSC-derived CECs for this cell therapy remain a challenge.Here we defined an adapted differentiation protocol to generate induced corneal endothelial cells (iCECs) consistently and efficiently from clinical-grade human embryonic stem cells (hESCs) with xeno-free medium and manufactured cryopreserved iCECs. Cells express high levels of typical CECs markers and exhibit transendothelial potential properties in vitro typical of iCECs. After rigorous quality control measures, cells meeting all release criteria were available for in vivo studies. We found that there was no overgrowth or tumorigenicity of grafts in immunodeficient mice. After grafting into rabbit models, the surviving iCECs ameliorated edema and recovered corneal opacity.Our work provides an efficient approach for generating iCECs and demonstrates the safety and efficacy of iCECs in disease modeling. Therefore, clinical-grade iCECs are a reliable source for future clinical treatment of corneal endothelial dysfunction.© 2023. The Author(s).