荧光介孔硅包覆铁氧化物神经干细胞标记研究

Research on fluorescent mesoporous silica-coated iron oxide labeled neural stem cell

目的以介孔硅包覆磁颗粒M-MS 50与SHU-555A对比,标记永生化神经干细胞C17.2。方法采用了核磁共振,普鲁士蓝染色,流式细胞仪,原子发射光谱,弛豫仪等一系列检测手段,测定细胞对磁颗粒的吞腹量,磁颗粒对细胞毒性的影响,及其在细胞内的代谢情况,选择细胞标记的最优条件。结果在神经干细胞对荧光介孔硅包覆铁氧化物的吞噬行为实验结果来看,随着孵育时间增长,SHU-555A和M-MS 50都更多的被C17.2神经干细胞所吞噬,这表现出细胞标记的时间依赖性。而M-MS 50在较短时间内便可以对细胞有一定的标记率。在标记神经干细胞的MRI和ICP-OES定量分析中可以得出,C17.2神经干细胞对SHU-555A和M-MS 50均表现出浓度依赖性和时间依赖性,SHU-555A和M-MS 50与C17.2细胞孵育,都存在标记率随时间梯度和浓度梯度的增加而提高,直到平衡这一现象。而通过荧光介孔硅包覆铁氧化物细胞内分布和代谢研究,发现随着标记时间的增长和标记浓度的增加,C17.2神经干细胞的标记率逐渐增加;磁颗粒有没有从细胞中代谢出来,并且在培养多长时间之后,细胞对M-MS 50的代谢达到平衡。结论介孔硅包覆氧化铁纳米颗粒M-MS 50在标记神经干细胞C17.2上相对SHU-555A有较明显的优势,而其多功能,特殊的介孔结构,又为以后进一步利用提供了更广阔的空间。M-MS 50能够有效地缩短周围质子的弛豫时间,提高弛豫信号强度,是一种非常优越的造影剂。

Objective To compare M-MS 50 with SHU-555 A of mesoporous silica-coated magnetic particles,label the immortalized neural stem cells C17. 2. Methods Swallowing abdominal volume of cells,the effect of magnetic particles on cell toxicity and its metabolism in cells were determined by a series of tests such as nuclear magnetic resonance,Prussian blue staining,flow cytometry,atomic emission spectrometry,relaxation instrument. The optimal conditions was screened according to the above-mentioned experimental result. Results The results of phagocytic behavior of neural stem cells on the fluorescent mesoporous silica-coated iron oxide showed that more and more SHU-555 A and M-MS 50 were phagocytosed by neural stem cells C17. 2with the incubation time increased,which illuminated the time dependence of cell labeling. While M-MS 50 could have a certain rate of cells labeling in a relatively short period of time. In MRI and ICP-OES,Phagocytization of neural stem cells C17. 2 on SHU-555 A and M-MS 50 showed a concentration and time dependent manner,SHU-555 A and M-MS 50 incubated with C17. 2 cells showed labeled rates increased with the increase of time and concentration gradients,until the balance. Intracellular distribution and metabolism research of,fluorescent mesoporous silica-coated iron oxide showed that neural stem cell C17. 2 labeling rate increased with the increase of time and concentration gradients,gradually. Conclusion Mesoporous silicacoated iron oxide nanoparticles of M-MS 50 has obvious advantages over SHU-555 A in labeling neural stem cells C17. 2,but its multifunctional,special mesoporous structure provides wider space for further use. M-MS 50 could effectively shorten the relaxation time of surrounding proton,improve the relaxation signal strength,which is a very excellent contrast agent.