基础医学与临床 ›› 2009, Vol. 29 ›› Issue (8): 859-862.

• 技术与方法 • 上一篇    下一篇

菲立磁胞内标记食蟹猴骨髓基质细胞

杨志军 罗永春 徐如祥 柯以铨 姜晓丹 蔡颖谦 杜谋选 邹雨汐   

  1. 北京军区总医院神经外科 北京军区总医院 北京军区总医院神经外科 南方医科大学珠江医院
  • 收稿日期:2009-01-23 修回日期:2009-04-03 出版日期:2009-08-20 发布日期:2009-08-20
  • 通讯作者: 徐如祥

Intracellular labeling of bone marrow stromal cells of Crab-eating Macaque with Feridex

Zhi-jun YANG, Yong-chun LUO, Ru-xiang XU, Yi-quan KE, Xiao-dan JIANG, Ying-qian CAI, Mou-xuan DU, Yu-xi ZOU   

  1. Department of Neurosurgery, General Hospital of Beijing Military District of Chinese PL Department of Neurosurgery, General Hospital of Beijing Military District of Chinese PL Zhujiang Hospital of Southern Medical University
  • Received:2009-01-23 Revised:2009-04-03 Online:2009-08-20 Published:2009-08-20
  • Contact: Ru-xiang XU,

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。

关键词: 骨髓基质细胞, 神经干细胞, 菲立磁, 超顺磁性氧化铁

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: Under the sterile condition, the crab-eating Macaque bone marrow stromal cells 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 microscope and immunocytochemistry. Results: BMSCs could be 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 under 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.

Key words: bone marrow stromal cells, neural stem cells, Feridex, superparamagnetie iron oxide