RiPS细胞用于脊髓损伤髓鞘修复的研究

项目名称： RiPS细胞用于脊髓损伤髓鞘修复的研究

项目编号： No.81200961

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 神经系统疾病、精神疾病

项目作者： 杨俊林

作者单位： 杭州师范大学

项目金额： 23万元

中文摘要： 脊髓损伤(SCI)后一项重要病理特征是少突胶质细胞凋亡后引起的脱髓鞘，少突胶质前体细胞(OPCs)移植修复髓鞘将是治疗SCI的重要途径之一。mRNA转染诱导的iPS细胞(RiPS细胞)因无基因组改变而成为OPCs安全的细胞来源。在本研究中，我们拟构建在人细胞中以附加体形式长期存在且能在OLIG2+细胞中特异表达GFP的载体:BAC-OG-Neo-OE，该BAC不会重组到人基因组而具有很好的安全性。将该BAC导入我们已经成功诱导的人RiPS细胞，建立RiPS细胞向OPCs分化的实验流程，在分化过程中利用OLIG2在OPCs特异表达的特点流式筛选GFP+的OPCs。将所获高纯度OPCs移植到SCI大鼠模型，研究其在损伤区域的存活、迁移、分化和参与髓鞘修复的情况。本研究将为利用安全的转基因技术从RiPS细胞定向分化中获得高纯度目的细胞开辟新的思路，为RiPS细胞用于SCI治疗积累重要的实验依据。

中文关键词： 脊髓损伤；髓鞘再生；细胞分化；少突胶质前体细胞；

英文摘要： One important pathophysiology and dysfunction of spinal cord injury (SCI) is the demyelination caused by the apoptosis of oligodendrocytes (OLs). Oligodendrocyte precursor cell (OPCs) transplantation for the remyelinating is an efficacious way to treat SCI. IPS cells induced by mRNA transfection (RiPS cells) provide a safe cell source for OPCs as no change in the genome. In the present study, we plan to build a bacterial artificial chromosome: BAC-OG-Neo-OE which can exist in human cells for a long time as a episome and can specifically express GFP in OLIG2-positive cells, and it will not recombine with the host genome, so it is a safe BAC for transgenic. This BAC will be tranfected into RiPS cells we have already induced, and then establish a protocol for the differentiation of RiPS cells to OPCs, and in the appropriate time of differentiation filter the GFP-positive OPCs by cell sorting utilizing the specific expression of OLIG2 in OPCs. The high-purity OPCs obtained by cell sorting are then transplanted into SCI rat model to study their survival, migration, differentiation, and remyelination in the injury area. This study will help to open up a new way to enrich high-purity interest cells from the directed differentiation of RiPS cells by this safe transgenic technology, and it will accumulate some important

英文关键词： Spinal cord injury；Remyelination；Cell differentiation；Oligodendrocyte precursor cells；