中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (19): 3436-3440.doi: 10.3969/j.issn.1673-8225.2010.19.004

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

超顺磁性氧化铁标记骨髓间充质干细胞体外诱导分化为神经细胞

王  凯，李建丁，张瑞平，徐隋意，汲  晶   

  1. 山西医科大学第一临床医学院放射科，山西省太原市  030001
  • 出版日期:2010-05-07 发布日期:2010-05-07
  • 通讯作者: 李健丁，教授，主任医师，博士生导师，山西医科大学第一临床医学院放射科，山西省太原市 030001 Cjr.lijianding@vip.163.com
  • 作者简介:王 凯，男，1984年生，山西省临汾市人，汉族，山西医科大学在读硕士，主要从事腹部影像诊断的研究。 lovewk2005@163.com

Differentiation of bone marrow mesenchymal stem cells labeled by superparamagnetic iron oxide into neural cells in vitro

Wang Kai, Li Jian-ding, Zhang Rui-ping, Xu Sui-yi, Ji Jing   

  1. Department of Radiology, First Clinical Medical College, Shanxi Medical University, Taiyuan   030001, Shanxi Province, China
  • Online:2010-05-07 Published:2010-05-07
  • Contact: Li Jian-ding, Professor, Chief physician, Doctoral supervisor, Department of Radiology, First Clinical Medical College, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China cjr.lijianding@vip.163.com
  • About author:Wang Kai★, Studying for master’s degree, Department of Radiology, First Clinical Medical College, Shanxi Medcal University, Taiyuan 030001, Shanxi Province, China lovewk2005@163.com

PDF

390

被引次数

8

摘要：

背景：一些研究者利用超顺磁性氧化铁对骨髓间充质干细胞进行标记，并利用MRI技术对标记细胞肝内、肾内移植后进行初步的活体示踪，但是，对于超顺磁性氧化铁标记骨髓间充质干细胞后，对其存活、增殖以及分化是否有影响研究较少。
目的：观察超顺磁性氧化铁标记骨髓间充质干细胞后，对其存活、增殖以及向神经细胞的分化是否有影响。
方法：使用超顺磁性氧化铁标记大鼠骨髓间充质干细胞，采用普鲁士蓝染色法鉴定其标记率、锥虫蓝染色法检测细胞活力、MTT法检测标记干细胞的细胞增殖活力、以1 mmol/L β-巯基乙醇及无血清DMEM培养液体外诱导标记细胞向神经细胞分化并用免疫组织化学奠定诱导后细胞、再次使用普鲁士蓝染色法鉴定神经细胞内的铁颗粒。
结果与结论：普鲁士蓝染色对干细胞的标记率接近100%，锥虫蓝染色显示标记细胞的存活率为97%；MTT法检测发现标记干细胞的增殖活力与未标记干细胞相比差异无显著性意义(P > 0.05)；以β-巯基乙醇诱导后大部分骨髓间充质干细胞分化为神经元样细胞、免疫组织化学阳性，再次普鲁士蓝染色显示铁颗粒位于神经细胞的细胞浆内。提示超顺磁性氧化铁标记骨髓间充质干细胞后，对于干细胞的存活、增殖以及向神经细胞的分化无影响。

关键词: 超顺磁性氧化铁, 骨髓间充质干细胞, 诱导分化, 神经细胞, 增殖

Abstract:

BACKGROUND: Some investigators used superparamagnetic iron oxide to label bone marrow mesenchymal stem cells (BMSCs). Magnetic resonance imaging was used to conduct primary living tracing of labeled cells in the liver and kidney following transplantation. However, there are a few reports addressing survival, proliferation and differentiation of superparamagnetic iron oxide-labeled BMSCs.
OBJECTIVE: To research whether there is any effect on cellular viability, proliferation and differentiation into neural cells of BMSCs following labeling with superparamagnetic iron oxide particles. 
METHODS: Superparamagnetic iron oxide was used to label rat BMSCs, and Prussian blue staining was used to identify labeling index. Trypan blue staining was employed to detect cell vitality. MTT assay was utilized to detect proliferation activity of labeled stem cells. Labeled stem cells were induced to differentiate into neural cells in vitro with 1 mmol/L β mercaptoethanol (BME) and serum-free DMEM. Immunohistochemistry was used to lay induced cells, and then Prussian blue staining was employed to identify the iron particles in neural cells again. 
RESULTS AND CONCLUSION: The stem cell labeling rate with Prussian blue staining was nearly 100%. The cell vitality rate with Trypan blue staining was 97%. MTT method detected that there was no significant difference for proliferation activity between labeled stem cells and those not labeled (P > 0.05). Majority of BMSCs differentiated into neuron-like cells after inducing by BME. Immunohistochemistry was positive. Prussian blue staining again showed that iron particles were in the nerve cell cytoplasm. These suggested that there is no effect on survival, proliferation and differentiation into neural cells of stem cells after it is labeled by superparamagnetic iron oxide.

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