人脐带组织富含间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs),是干细胞研究理想的种子来源,如何从脐带组织中分离间充质干细胞及运用影像技术示踪干细胞生物学行为是当前研究的热点。该实验应用组织块贴壁法从足月孕妇脐带组织中分离纯化间充质干细胞并进行鉴定,结果为hUCMSCs。进一步应用磷酸钙、Effectene、脂质体2000三种转染试剂分别介导Gd-DTPA标记hUCMSCs,通过MRI(magnetic resonance imaging)检测钆喷酸葡胺(Gd-DTPA)标记细胞信号强度变化及细胞内钆离子浓度的测定评价三种转染试剂的转染能力,最终发现在三种转染试剂中,Effectene介导Gd-DTPA标记hUCMSCs效果最佳,为Gd-DTPA标记干细胞体外MR成像奠定了实验基础。
Human umbilical cord tissue contains plenty of mesenchymal stem cells, which is the ideal stem cell seed source. How to isolate mesenchymal stem cells from umbilical cord and trace the biological behavior of stem cells by image techniques are current research focuses. The hUCMSCs from full-term pregnant women's umbilical cord tissues were separated, purified and identified in this research. Furthermore, Gd-DTPA was transfected into mesenchymal stem cells through calcium phosphate, Effectene and Lipofectamine 2000. The change of labeled cell's signal strength was detected by MR and the gadolinium ion concentration intra celluar were determined by inductively coupled plasma atomic emission spectrometry for evaluation of the transfection capacity of three reagents. In conclusion, the method of Effectene mediated Gd-DTPA marker hUCMSCs is best and this study provides the experimental foundation for Gd-DTPA labeled stem cells in vitro MR.