【摘要】 目的探索利用菲立磁和转染试剂体外磁性标记食蟹猴骨髓基质细胞(BMSCs)方法的可行性。方法无菌条件下取食蟹猴骨髓,梯度密度离心法分离获取BMSCs,使用菲立磁-多聚赖氨酸复合物(FE-PLL)标记BMSCs,普鲁士蓝染色、电镜和台盼蓝排除实验等方法鉴定FE-PLL标记食蟹猴BMSCs的效率和细胞的活力,倒置相差显微镜和免疫细胞化学方法检测BMSCs的增殖和分化能力。结果菲立磁可以高效率地标记BMSCs,标记效率在99%左右。光镜和电镜下BMSCs胞质内分别可见细小的蓝色铁颗粒和许多包裹铁颗粒的囊泡。FE-PLL标记对BMSCs的活力、增殖和分化等能力没有明显的影响。结论菲立磁可以用来体外标记食蟹猴BMSCs。更多还原

【Abstract】 Objective To explore the feasibility of protocols using Feridex and transfection agents for in vitro magnetic labeling of bone marrow stromal cells (BMSCs).Methods The bone marrow stromal cells of crab-eating Macaque were harvested by means of density gradient centrifugation following a bone puncture. Feridex-poly-l-lysine (FE-PLL) complexes were used to magnetically label BMSCs. The efficiency and cellular viability of FE-PLL labeled BMSCs were evaluated by Prussian blue staining,electron microscopy,and trypan blue dye exclusion test. The proliferation and differentiation ability of FE-PLL labeling BMSCs were also investigated by inverted phase contrast microscopy and immunocytochemistry. Results BMSCs were effectively labeled and labeling efficiency were around 99%. Numerous blue-stained fine particles and numerous vesicles filled with the electron-dense magnetic iron particles could be found in the cytoplasm of FE-PLL labeled BMSCs in optical microscopy and transmission electron microscopy respectively. Cell viability,proliferation and differentiation ability of labeled cell were not affected by endosomal incorporation of Feridex nanoparticles. Conclusion The Feridex might be used to label BMSCs of crab-eating Macaque.更多还原