关键词：
壳聚糖薄膜;球体培养;间充质干细胞;迁徙趋化
摘 要：
基于细胞实验研究壳聚糖(chitosan,CS)薄膜成球培养技术对间充质干细胞(mesenchymal stem cells,MSCs)迁徙趋化特性的影响.从脐带组织中分离原代MSCs采取CS成球法培养,以常规贴壁培养MSCs作为对照,72 h后收集两组细胞分别进行划痕实验、Tranthwell迁徙实验观察并拍照记录,RT-PCR方法检测两种培养方式中MSCs迁徙相关基因表达水平的差异.研究结果显示,相较常规贴壁培养方式,CS培养组MSCs体外迁徙趋化能力增强,差异具有显著统计学意义(P<0.01);CS成球培养组MSCs中CXCR4、CXCR7、MCP-1、MMP-1、MMP-2、MMP-9、TIMP-2等迁徙相关基因表达均明显上调(P<0.01).实验表明CS成球培养可显著促进MSCs的迁移趋化特性.
作 者：
LI Huajie;XU Xiongfeng;QIU Bo;Department of Orthopedics,Renmin Hospital of Wuhan University;
单 位：
LI Huajie%XU Xiongfeng%QIU Bo%Department of Orthopedics,Renmin Hospital of Wuhan University
关键词：
chitosan membranes;;spheroid formation;;mesenchymal stem cells;;migration and chemotaxis
摘 要：
To investigate the effects of chitosan-mediated spheroid formation on the migration and chemotaxis of mesenchymal stem cells( MSCs). The original MSCs were extracted from umbilical cord tissue,and were cultured on chitosan( CS) membranes,compared with these on trational adherent substrate. MSCs of two groups were collected after 72 hours. Migration rate and chemotactic effect of MSCs were measured by wound healing assay and tranthwell migration assay,observed and screened the cell morphology. The expression levels of genes related to MSCs migration and chemotaxis in two cultures were further detected by real-time PCR( RT-PCR). The results showed that,compared with the MSCs derived on adherent substrate,there was a significant statistical difference( P<0.01) that MSC spheroids derived on CS have more active migration and chemotaxis in vitro.And the expression level of migration genes,such as CXCR4,CXCR7,MCP-1,MMP-1,MMP-2,MMP-9 and TIMP-2,was significantly up-regulated in the MSC spheroids derived on CS( P<0.01). The experimental results showed that chitosan-mediated spheroid formation could enhanced the activities of migration and chemotaxis of MSCs.
