人脐血间充质干细胞成骨潜能亚群的特异性分子标志

项目名称： 人脐血间充质干细胞成骨潜能亚群的特异性分子标志

项目编号： No.30800232

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 刘广鹏

作者单位： 上海交通大学

项目金额： 20万元

中文摘要： 如何获得数量充足、功能良好且具有成骨分化潜能的种子细胞，一直是困扰骨组织工程基础研究与产业化发展的主要问题。具有多向分化潜能的人脐血间充质干细胞（UCB-MSCs）来源充足，细胞含量高，有望成为骨组织工程的种子细胞。然而，目前常用分离方法获得的UCB-MSCs中含有大量混杂细胞，严重影响了UCB-MSCs诱导成骨的成功率和均质性，因此，必须对其进行分选纯化。本项目拟通过单细胞克隆培养及成骨定向诱导鉴定的方法，将UCB-MSCs中具有克隆形成能力的细胞分为成骨分化潜能和非成骨分化潜能的两类克隆群，再经过基因组学分析全面比较两类克隆细胞膜表面蛋白表达谱的差异，去同存异，从中筛选出与成骨分化潜能相关的膜蛋白，进一步通过流式细胞分析、免疫磁珠分选后成骨诱导，最终确定具有成骨分化潜能细胞亚群的特异性分子标志，为建立稳定有效的UCB-MSCs成骨潜能亚群分选与纯化技术提供技术基础和科学依据。

中文关键词： 组织工程；脐带血；间充质干细胞；成骨潜能; 细胞分选

英文摘要： To harvest a sufficient amount of seed cells with osteogenic potential is the main purpose for bone tissue engineering. Mesenchymal stem cells derived from human umbilical cord blood (UCB-MSCs), with the characteristics of enough supply and high cell production, may represent a new source of seed cells for tissue engineering. However, the UCB-MSCs obtained with present separating methods always contain a large number of undesired, mixed cell types, resulting in the failure of successful osteogenic induction in vitro. The objective of this study is to investigate the possibility of sorting and selecting UCB-MSCs with osteogenic potential. With single-cell-cloning culturing and osteogenic-induction identification, UCB-MSCs which can form clones would be divided into two types: with or without the potential of osteogenic differentiation. By comparing the difference of surface protein expression between these two types of cells with gene analysis, certain surface proteins related to the osteogenic potential would be seleted. Then, such specific molecular markers could be identified by osteogenic differentiation after flow cytometric analysis and immunological bead sorting. The results of this project are expected to provide basis for establishing stable and effective techniques to sort UCB-MSCs with osteogenic potential specifically.

英文关键词： tissue engineering; umbilical cord blood; mesenchymal stem cells; osteogenic potential; cell sorting