角膜内皮功能失代偿会导致角膜混浊和视力下降，而成人角膜内皮细胞增殖能力有限，因此，为解决这一难题需要寻找新的替代细胞来源。人多能干细胞是治疗致盲性眼病的理想种子细胞。我们发现，维甲酸（retinoid acid，RA）诱导后的人胚胎干细胞能够在ROCK抑制剂的调控下进一步分化为角膜内皮细胞，表达Na+-K+-ATPase，具有体内泵水功能，细胞形态和性质特征与人角膜内皮细胞相似。在此基础上，本课题拟进一步开展以下研究：（1）建立RA联合ROCK抑制剂诱导人多能干细胞定向分化为功能性角膜内皮细胞的实验体系，对分化角膜内皮细胞进行形态学、相关基因、蛋白表达及体内外功能鉴定；（2）检测抑制或激活Rho-ROCK信号通路时角膜内皮功能蛋白表达情况，探讨ROCK抑制剂在调控功能性角膜内皮细胞分化中的作用机制。为临床角膜内皮移植提供种子细胞来源，为角膜内皮功能失代偿的治疗提供新的思路和实验依据。

Corneal endothelial dysfunction could cause corneal edema and even vision loss.However, adult human corneal endothelial cells show limited proliferative capacity both in vivo and in vitro.Human pluripotent stem cells (hPSCs) have been confirmed as an ideal cell source of blinding diseases therapy. In our previous study, we found that hPSCs could be induced into functioinal corneal endothelial cells by ROCK inhibitor after treat with retinoid acid (RA). The differentiated cells with function of water pump in vivo showed similar morphology and typical gene expressions with human corneal endothelial cells. In this project, we will further examine the characteristics and functions of hPSC-derived corneal endothelial cells in vitro and in vivo, demonstrate the mechanism of ROCK inhibitor during the differentiation process of functional corneal endothelial cells. Our findings will not only reveal the molecular mechanisms of corneal endothelial differentiation, but also supply enough source cells for the clinical therapy of corneal endothelial dysfunction.