近年来，胰岛样细胞替代疗法被认为是一种很有希望治愈糖尿病的途径。我们的前期工作初步探明Pdx-1基因在转分化BMSCs为胰岛样细胞中发挥重要作用，但所获胰岛样细胞的超微结构和功能尚不理想。如何优化胰岛样细胞的诱导分化条件并探明其中机制是亟待解决的难题。本项目将在前期研究工作的基础上，以BMSCs做为靶细胞，采用可后期直接应用于临床的三个抗不同抗生素的质粒，1）筛选能稳定高效表达Pdx-1, Ngn3 和MafA基因的转化细胞株，明确这三个转录因子的特定组合影响包括胰岛素在内的相关基因的表达情况； 2）建立在GLP-1 和gastrin的诱导下能有效分泌胰岛素的工程细胞株，探讨GLP-1和gastrin进一步促进胰岛样细胞增殖的分子机制；3）鼠肾被膜移植，疗效评定及免疫组织化学分析。本项目的研究成果将建立完善的胰岛样细胞分化技术，为临床推广奠定理论及实践基础，具有重要的临床应用价值及社会效益

In recent years, insulin-producing cells substitution therapy is considered to be a promising treatment to cure diabetes. Our preliminary studies initially proved the effects and mechanisms of Pdx-1 on transdifferentiation of BMSCs into insulin-producing cells, but the ultrastructure and function of insulin-producing cells which we obtained are not ideal. In this study the optimized method suitable for induction and differentiation conditions of insulin-producing cells will be screen and investigated.In this project based on the previous studies, the three kinds of plasmids against different antibiotics, which can be directly applied to clinical practise, will be transfected into the target cells BMSCs. Subsequently, the following experimental studies will be carried out: 1)screening the transfected cell lines which can efficiently and stably express the Pdx-1, Ngn3 and MafA genes, and investigating the specific combined effects of the three transcription factors on the related gene expression, including the insulin gene. 2) establishing GLP-1 and gastrin induced insulin-secreting engineering cells, and exploring the molecular mechanisms of GLP-1 and gastrin induced proliferation of insulin-producing cells.3) the efficacy evaluation and immunohistochemical analysis of the rat renal capsule transplantation. This project will help us to establish consummate differentiation technology of insulin-producing cells and lay a foundation of clinical theory and practice.