骨髓间充质干细胞来源和成纤维细胞来源的iPS细胞向功能性成熟红细胞分化潜能差异及其机理研究

项目名称： 骨髓间充质干细胞来源和成纤维细胞来源的iPS细胞向功能性成熟红细胞分化潜能差异及其机理研究

项目编号： No.81200338

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 徐玉林

作者单位： 浙江大学

项目金额： 23万元

中文摘要： 将诱导性多能干细胞（iPS）分化为功能性红细胞是目前国际研究热点。在前期工作中，我们建立了高效诱导体细胞重编程为iPS细胞及诱导iPS细胞分化为造血干细胞的技术平台。我们发现骨髓间充质干细胞（MSC）来源较成纤维细胞（Fib）来源的iPS细胞分化为造血细胞效率更高，具体机理还不清楚。本课题拟研究MSC来源和Fib来源的iPS细胞分化为功能性成熟红细胞的潜能、潜能差异及相关机理。通过检测整个诱导过程中细胞表面标记、目的细胞获得率、各种珠蛋白的表达及比例、携氧能力和Bohr效应等明确两种细胞来源的iPS细胞分化潜能及潜能差异；同时研究造血相关基因如SCF、GATA-1等启动子甲基化情况、基因表达、蛋白表达及表达量等明确差异机理。本研究为探索iPS细胞分化效率是否受到它们起源的细胞类型的表观遗传记忆影响提供依据，为建立高效红系诱导体系提供合适的细胞来源，也为红系疾病机理研究等提供理想的细胞模型。

中文关键词： 诱导性多能干细胞；骨髓间充质干细胞；成纤维细胞；分化体系；造血细胞

英文摘要： Induced pluripotent stem (iPS) cells are a great breakthrough in bioscience and it provides an unprecedented platform to generate functional red blood cells which is a useful treatment in clinical application. Based on the previous work of the efficient generation of iPS cells and the efficient commitment iPS cells to functional CD34+ progenitors, we found that human bone marrow mesenchymal stem cell-derived iPS cells (MSC-iPS) are more efficient than human fibroblast (Fib)-derived ones in producing CD34+ progenitors.However, the mechanism is unclear. The goal of the present study is to assess the induction capacity difference of MSC- and Fib-derived iPS cells into functionally mature RBC and its mechanism. The former is determined by analysis of several indicators as the expression of cell markers, the efficiency of red blood cells, the expression of types of hemoglobins and their efficiency during the induction. At the same time, the mechanism is studied with the focus in some hematopoietic factors as SCF and GATA-1 for their promoter methylation, the gene expression, the protein expression and the expression percentage. The study provides proofs whether the epigenetic memory of their cell-derivation lineages of induced pluripotent stem cells will be preferential for iPS cell lineage-specific differentiation

英文关键词： induced pluripotent stem cells；Mesenchymal stem cells；Fibroblasts；differentiation systems；hematopoietic cells