The development of basic technologies for the cellular therapy of corneal epithelial cells by the regulation of cellular senescence and epigenetic changes

点击翻译按钮获取中文摘要

In this study, in order to establish the method to transplant corneal epithelial cells which are amplified ex vivo from the limited number of autologous corneal epithelial cells, we investigated the cellular senescence and epigenetic changes of the cultured corneal epithelial cells as well as the possible countermeasures against these culture-associated drawbacks. We have found that the expression of the keratin 12 gene significantly decreased rapidly after the cell dissociation of the corneal epithelial cells, which was found to be due to the activation of several signaling pathways such as MAPK accompanied with the epigenetic changes of the keratin 12 promoter. The proliferation of the corneal epithelial cell was found to be significantly enhanced by the introduction of SV40 large T antigen which is known to inhibit p53 and Rb tumor- suppressive pathways. Quite interestingly, when corneal epithelial stem cells were transfected with the SV40 large T antigen, the endogenous hTERT gene was found to be activated, which is thought to be the risk to lead the tumorigenic transformation of these cells.