Yu Juan, Yu Nianye, Tian Yao, Fang Yifan, An Bin, Feng Guihai, Wu Jun, Wang Liu, Hao Jie, Wang Liqiang, Zhou Qi, Li Wei, Wang Yukai, Hu Baoyang
Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China.
Institute of Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China.
Cell Biosci. 2023 Nov 6;13(1):201. doi: 10.1186/s13578-023-01145-w.
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.
人类多能干细胞（hPSCs）的研究在基于细胞的再生医学领域取得了巨大进展。角膜内皮功能障碍与角膜内皮细胞（CECs）的丢失和退化相关，使得细胞替代成为一种有前景的治疗策略。然而，对于这种细胞疗法，对hPSC来源的CECs进行全面的临床前评估仍然是一项挑战。
在此，我们定义了一种适应性分化方案，以使用无动物成分培养基从临床级人类胚胎干细胞（hESCs）中持续且高效地生成诱导角膜内皮细胞（iCECs），并制备了冷冻保存的iCECs。细胞表达高水平的典型CECs标志物，并在体外表现出iCECs典型的跨内皮电位特性。经过严格的质量控制措施后，符合所有放行标准的细胞可用于体内研究。我们发现免疫缺陷小鼠体内的移植物没有过度生长或致瘤性。将其移植到兔模型中后，存活的iCECs减轻了水肿并恢复了角膜混浊。
我们的工作为生成iCECs提供了一种有效的方法，并证明了iCECs在疾病建模中的安全性和有效性。因此，临床级iCECs是未来角膜内皮功能障碍临床治疗的可靠来源。
