菲立磁细胞内标记兔骨髓基质细胞的初步观察

Preliminary study on intracellular labeling of neural stem cells derived from rabbit bone marrow with Ferumoxides

目的:探讨利用菲立磁(Feridex)和转染试剂体外标记兔骨髓基质细胞的可行性,为临床上应用磁共振成像(MRI)追踪标记细胞奠定基础。方法:9只2月龄新西兰大白兔无菌条件下行股骨穿刺取骨髓,梯度密度离心法分离获取兔骨髓基质细胞,用菲立磁-多聚赖氨酸复合物(FE-PLL)标记骨髓基质细胞,采用普鲁士蓝染色和台盼蓝拒染实验等方法鉴定FE-PLL标记兔骨髓基质细胞的效率和细胞的活力,并在增殖条件下用改良SABC法进行抗单克隆神经元特异性烯醇化酶(NSE)免疫组织化学染色(DAB-H2O2棕色法呈色)和普鲁士蓝(Prussian blue)染色,对FE-PLL标记骨髓基质细胞的增殖和分化能力进行评估。结果:菲立磁可以高效率地标记兔骨髓基质细胞,标记效率在99%左右。普鲁士蓝染色显示FE-PLL标记骨髓基质细胞胞质内出现细小的蓝色铁颗粒。标记的骨髓基质细胞与正常未标记细胞相比较,细胞的活力、增殖和分化等能力没有明显的差异。结论:菲立磁可以用来体外标记骨髓基质细胞,标记后对骨髓基质细胞活力、增殖和分化能力无明显影响。

Objective：To explore the feasibility of protocols using Feridex and transfection agents for in vivo magnetic labeling of bone marrow stromal cells（BMSC） and provided the evidence for the tracing of labeled cells clinically by magnetic resonance imaging （MRI） in the future.Method：Under the sterile condition,the rabbit bone marrow was aspirated from the femur,and BMSC were harvested by means of density gradient centrifugation . and labeled magnetically by feridex-poly-l-lysine（FE-PLL） complexe.The efficiency and cellular viability of FE-PLL labeled BMSC were evaluated by Prussian blue staining,and trypan blue dye exclu- sion test.The proliferation and differentiation ability of FE-PLL labeling BMSC were also investigated.Result： BMSC could be effectively labeled and labeling efficiency was about 99%.Prussian blue staining showed nu- merous blue stained fine particles in the cytoplasm of FE-PLL labeled BMSCs.The viability,proliferation and differentiation ability of labeled BMSC were not affected by endosomal incorporation of Feridex nanoparticles compared to control unlabeled cells.Conclusion：Feridex can be used to label BMSC,and viability,proliferation and differentiation ability are not affected by endosomal incorporation of Feridex nanoparticles.