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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 investigate the feasibility of in vitro labeling of rabbit bone marrow stromal cells with Feridex and transfection reagents, and lay a foundation for the clinical application of magnetic resonance imaging (MRI) for tracing labeled cells. Methods: Nine rabbits of 2 months old New Zealand white rabbits were subjected to femoral bone puncture under sterile conditions. Rabbit bone marrow stromal cells were isolated by gradient density centrifugation. Marrow stromal cells were labeled with Fe-Poly-Lysine Complex (FE-PLL) Cells were stained with Prussian blue and trypan blue exclusion to identify the efficiency and viability of FE-PLL labeled rabbit bone marrow stromal cells. Anti-monoclonal neuron-specific enolase Enzyme-linked immunosorbent assay (NSE) immunohistochemical staining (DAB-H2O2 brown staining) and Prussian blue staining were used to assess the proliferation and differentiation of FE-PLL labeled bone marrow stromal cells. Results: Philippine magnetic beads can mark rabbit bone marrow stromal cells efficiently, labeling efficiency of about 99%. Prussian blue staining showed the presence of fine blue iron particles in the cytoplasm of FE-PLL labeled bone marrow stromal cells. Markers of labeled stromal cells compared with normal unlabeled cells, cell viability, proliferation and differentiation ability no significant difference. Conclusion: Feridex could be used to label bone marrow stromal cells in vitro, and markedly had no effect on the viability, proliferation and differentiation of bone marrow stromal cells.