我国约有120万角膜内皮盲患者，需通过角膜内皮移植进行治疗。但角膜供体严重短缺及术后免疫排斥等制约了该方法的开展。构建人组织工程角膜内皮(TECE)有望为此类患者带来福音,但如何获取TECE的种子细胞一直是个难题。在课题组成员前期工作的基础上，本项目拟用诱导多潜能干细胞（iPSCs）技术制备TECE所需的自体角膜内皮细胞（CEC）。我们首次提出通过电穿孔导入多能细胞抽提物的方法，利用与CEC来源相近的体细胞制备无病毒的稳定的iPSCs,并对所制备的iPSCs进行形态学鉴定、多潜能特性、体内外分化能力及核型分析等鉴定；通过共培养及因子诱导等方法将所制备的iPSCs分化为角膜内皮细胞，并对其进行形态学、组织化学的鉴定及体内外生理功能的评估。TECE的成功将解决临床角膜材料的供需矛盾，并降低术后排斥的风险。本研究将为进一步研究角膜内皮细胞胚胎发育过程中各种细胞因子的作用及调控机制提供理论基础。

Lacking of donor corneas is a major problem in China. Tissue engineered cornea will effectively alleviate the pressure of this. Our team members have developed a suitable cornea carrier system, and set up the platform for generation induced pluripotent stem cells (iPSC). In this project, We hope to build tissue-engineered corneal endothelium using iPSCs generated from patient’s own cell.We plan to firstly generate iPSCs from both human skin fibroblasts and corneal stromal cells using several virus-free method, and then differentiate iPSCs into corneal endothelial cells. By doing there , we will find out the best somatic cell model for the treatment of corneal blindness，as well as provide the molecular basis for targeted iPSC differentiation. With tissue engineering technology, The success of this research will eliminate the dependence of cornea donor and reduce the risk of rejection and second transplantion. It will also be beneficial for exploring the functions and regulatory mechanism of corneal endothelial cell differentiation during embryonic development.