超顺磁性氧化铁Resovist标记神经干细胞的实验研究

Experimental Study on Resovist Labeled Neural Stem Cells

目的研究超顺磁性氧化铁(superparamgneticironoxides.SPIOs)标记神经干细胞及其生物学特性。方法:神经干细胞培养、传代和诱导分化;Resovist(一种SPIOs)标记神经干细胞,制备磁标记神经干细胞;利用免疫细胞化学、透射电镜和Prussianblue染色等方法对磁标记神经干细胞生物学特性进行研究。结果:在原代及传代细胞中有Nestin阳性细胞即神经干细胞。血清诱导下,神经干细胞可分化为GFAP、NF200阳性细胞。Resovist与神经干细胞共同孵育后,透射电镜及Prussianblue染色显示胞浆中含有铁颗粒,Resovist也可以随细胞的分裂增殖而传到子代细胞中。随Resovist浓度的增高(5.6μg/ml-11.2μg/ml),Resovist对神经干细胞存活、分化能力的影响无显著性差异(P>0.05)。当Resovist的浓度大于22.4μg/ml时,Resovist影响其存活和分化(P<0.05)。结论:本实验利用Resovist作为磁标记探针,对神经干细胞进行成功磁标记。为进一步利用核磁共振(MRI)对神经干细胞活体追踪奠定实验基础。

Objective To study the biology characteristic of superparamgnetic iron oxides （SPIOs） labeled neural stem cells（NSCs）. Methods The culture, passage and differentiation of NSCs from rat brain were carried out. Resovist labeled NSCs were use to be magnetic labeled NSCs. With immunocytochemistry,transmission electron microscopy（ TEM ） and Prussian blue staining, the growth, differentiation and other biology characteristics were examined. Results Nestin positive cells were found in the culture and dauhgter clones. NSCs were differentiated into positive GFAP and NF200 cells in serum culture. When NSCs were incubated with Resovist, the iron particles appeared within the cells （non-nucleus） and also existed in daughterclones. With the concentration of Resovist increased（5.6μg/ml-11. 2 μg/ml）, there was no obvious difference of the growth and differentiation of NSCs（P〉0.05）. When the concentration of Resovist exceeded 22.4μg/ml,there existed an obvious difference （P〈0.05）. Conclusion NSCs are successfully labeled with Resovist that is a kind of SPIOs to be magnetic labeled NSCs. That is the base of tracking magnetic labeled NSCs in vivo with MRI.