中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 6068-6072.doi: 10.3969/j.issn.1673-8225.2010.32.040

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

菲立磁胞内标记食蟹猴骨髓基质细胞

陈 霞1，杨志军2，罗永春2，蔡颖谦3，杜谋选3，邹雨汐3   

  1. 1解放军324医院儿科，重庆市  400020；2解放军北京军区总医院神经外科，北京市  100700；3南方医科大学珠江医院神经外科，广东省广州市  510282
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 杨志军，副主任医师，解放军北京军区总医院神经外科，北京市 100700 zhijunyangfmmu@yahoo.com.cn
  • 作者简介:陈 霞★，女，1975年生，重庆市人，汉族，2006年解放军第三军医大学毕业，硕士，主治医师，主要从事儿科新生儿学的研究。 06chenxia@163.com
  • 基金资助:

    解放军成都军区医学科研计划课题(MB07005)。

Feridex labeling of bone marrow stromal cells of crab-eating macaque

Chen Xia1, Yang Zhi-jun2, Luo Yong-chun2, Cai Ying-qian3, Du Mou-xuan3, Zou Yu-xi3   

  1. 1 Pediatric Department, the 324 Hospital of Chinese PLA, Chongqing   400020, China; 2 Department of Neurosurgery, General Hospital of Beijing Military Area Command of Chinese PLA, Beijing   100700, China; 3 Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou   510282, Guangdong Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Yang Zhi-jun, Associate chief physician, Department of Neurosurgery, General Hospital of Beijing Military Area Command of Chinese PLA, Beijing 100700, China zhijunyangfmmu@yahoo.com.cn
  • About author:Chen Xia★, Master, Attending physician, Pediatric Department, the 324 Hospital of Chinese PLA, Chongqing 400020, China 06chenxia@163.com
  • Supported by:

     Medical Scientific Research Plan of Chengdu Military Area Command of Chinese PLA, No. MB07005*

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摘要：

背景: 有关利用菲立磁体外细胞标记研究选择的动物主要是啮齿类动物，而研究灵长类动物食蟹猴的报道目前仍是空白。
目的：观察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骨髓基质细胞，使用菲立磁-多聚赖氨酸复合物标记骨髓基质细胞，普鲁士蓝染色、电镜和锥虫蓝排除实验等方法鉴定菲立磁-多聚赖氨酸复合物标记食蟹猴骨髓基质细胞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骨髓基质细胞的增殖和分化能力。
结果与结论：菲立磁可以高效率地标记骨髓基质细胞，标记效率在99%左右。光镜和电镜下骨髓基质细胞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。菲立磁-多聚赖氨酸复合物标记对骨髓基质细胞的活力、增殖和分化等能力没有明显影响。提示菲立磁可以用来体外标记食蟹猴骨髓基质细胞。

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:

BACKGROUND: Studies regarding Feridex in vitro cell labeling are mainly in rodents, while little information is known on primate crab-eating macaque.
OBJECTIVE: To explore the feasibility of protocols using Feridex and transfection agents for in vitro magnetic labeling of bone marrow stromal cells (BMSCs) in crab-eating macaque.
METHODS: Under the sterile condition, the crab-eating macaque BMSCs were obtained by means of density gradient centrifugation following a bone puncture. Feridex-poly-l-lysine complexes were used to magnetically label BMSCs. The efficiency and cellular viability of Feridex-poly-l-lysine labeled BMSCs were evaluated by Prussian blue staining, electron microscopy, and trypan blue dye exclusion test. The proliferation and differentiation ability of Feridex-poly-l-lysine labeling BMSCs were also investigated by inverted phase contrast microscope and immunocytochemistry.
RESULTS AND CONCLUSION: BMSCs could be effectively labeled by Feridex and labeling efficiency was around 99%. Tiny blue stained fine particles and numerous vesicles coated with the electron-dense magnetic iron particles could be found in the cytoplasm of Feridex-poly-l-lysine labeled BMSCs under optical microscopy and transmission electron microscopy respectively. Cell viability, proliferation and differentiation ability of labeled BMSCs were not affected by Feridex-poly-l-lysine labeling. Results indicated that Feridex might be used to label BMSCs of crab-eating macaque.

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