角膜内皮功能失代偿导致角膜混浊和视力下降是临床治疗的难题，角膜内皮细胞增殖能力非常有限，寻找自体来源丰富的多潜能干细胞移植是解决这一难题的有效方法。我们在前期研究中初步证实了骨髓内皮祖细胞在体外可诱导分化为角膜内皮样细胞并在猫眼中具有一定的修复角膜内皮缺损的能力，但其诱导条件和治疗效果尚需进一步探讨。为此我们拟结合niche微环境调控干细胞分化的理论，以及Rho/ROCK通路与细胞屏障作用的关系，深入研究：免疫磁珠提纯内皮祖细胞，通过角膜内皮和晶状体上皮细胞共培养体外诱导，前房注射诱导后的祖细胞至角膜内皮微环境中，探讨体外诱导加体内微环境作用后其分化再生修复角膜内皮的能力，同时加入促进形成细胞屏障作用的Rho/ROCK通路抑制剂，以达到完全修复内皮层的作用。研究结果有望真正实现自体干细胞移植代替角膜内皮细胞，不仅来源充足，而且避免了免疫排斥反应，为临床治疗角膜内皮失代偿疾病提供新的方法。

Corneal endothelial dysfunction caused by various diseases is very difficult to treat in clinics, resulting in corneal opacity and visual loss. The proliferation of human corneal endothelial cells (HCEC) is limited both in vivo and in vitro. Therefore, the key solution to the problem would be replacing the excessively lost HCECs with transplanted pluripotent stem cells which have rich sources. In the preliminary studies, we found that bone marrow-derived endothelial progenitor cells can be induced into corneal endothelial like cells through co-culture system with HCECs. And based on the theory that cell niche controls the differentiation of stem cells, Rho/ROCK signal is contribute to cell barrier.We intend to undertake the following studies: Using immunomagnetic beads to label and purify stem cells. Endothelial progenitor cells will be induced in vitro by co-culture with CEC and epithelial cells of lens.Then the induced progenitor cells will be injected into anterior chamber of rabbits, and then be oriented to attach on the surface of corneal Descemet's membrane by magnetic field outside. Thus, the feasibility of endothelial progenitor cells being induced to corneal endothelial cells in specific niche can be studied. At the same time, the role and mechanism of Rho/ROCK signaling pathway on enhancing the barrier of endothelial progenitor cells will be studied as well. The success of the study is expected to seek a real alternative of autologous stem cells to replace corneal endothelial cells. The advantages not only lie in the sufficient source of cells, but also that immune rejection can be avoided, and this method can overcome the limitations of biological materials. The results of this study can also provide new ideas and method for the clinical treatment of corneal endothelial dysfunction.