中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7420-7424.doi: 10.3969/j.issn.1673-8225.2010.40.003

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

人骨髓间质干细胞的体外菲立磁标记

钟跃思，林继宗，柯应平，汤照峰，徐见亮，林  楠，许瑞云，邓美海　   

  1. 中山大学附属第三医院肝胆外科，广东省广州市 510630
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 通讯作者: 邓美海，博士，主任医师，硕士生导师，中山大学附属第三医院肝胆外科，广东省广州市 510630 drdmh@medmail.com.cn
  • 作者简介:钟跃思☆，男，1979年生，广东省揭阳市人，汉族，2009年中山大学毕业，博士，医师，主要从事肝胆外科及干细胞治疗肝纤维化方面的研究。zhongyuesi@gmail.com
  • 基金资助:

    广东省自然科学基金(8151008901 000112)，课题名称：基因转染的骨髓间质干细胞治疗肝纤维化的实验研究。教育部博士学科点专项科研基金(20070558259)，课题名称：骨髓间质干细胞对肝星状细胞的调控作用及机理研究。

Human bone marrow mesenchymal stem cells labeled with Feridex in vitro

Zhong Yue-si, Lin Ji-zong, Ke Ying-ping, Tang Zhao-feng, Xu Jian-liang, Lin Nan, Xu Rui-yun, Deng Mei-hai　   

  1. Department of Hepatobiliary Surgery, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China　
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Deng Mei-hai, Doctor, Chief physician, Master’s supervisor, Department of Hepatobiliary Surgery, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China drdmh@medmail. com.cn
  • About author:Zhong Yue-si☆, Doctor, Physician, Department of Hepatobiliary Surgery, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China zhongyuesi@gmail.com
  • Supported by:

     the Natural Science Foundation of Guangdong Province, No.8151008901000112*; Specialized Research Fund for the Doctoral Program, Ministry of Education, No. 20070558259*

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摘要：

背景：常规干细胞示踪的检测方法，需取实验动物组织进行检查，缺乏示踪的动态检测和无创性。
目的：通过体外菲立磁标记骨髓间质干细胞并进行磁共振成像扫描，明确不同浓度菲立磁对人骨髓间质干细胞细胞活性的影响和检测菲立磁标记的最佳浓度，并筛选出适合人骨髓间质干细胞磁共振成像扫描的成像序列。
方法：使用不同质量浓度菲立磁(100，50，25，12.5 mg/L)与生长状态良好第3代人骨髓间质干细胞孵育24~48 h，另设未进行标记的第3代细胞为空白对照组。两组细胞进行四甲基偶氮唑蓝法检测增殖生长情况并描绘生长曲线。细胞普鲁士蓝染色鉴定人骨髓间质干细胞的标记情况。进行磁共振成像检测，确定最佳磁共振成像序列。
结果与结论：质量浓度≤25 mg/L的菲立磁标记对人骨髓间质干细胞的增殖能力不会产生影响，并且以25 mg/L的标记效果最佳。而≥50 mg/L的菲立磁标记则明显抑制人骨髓间质干细胞的增殖。磁共振成像扫描显示菲立磁标记24，48 h或子代的人骨髓间质干细胞标本T1WI、T2WI以及T2*WI 3个序列上均可见信号降低，以T2*WI变化最为明显。证实菲立磁可用于体外标记人骨髓间质干细胞连续性研究。菲立磁标记对人骨髓间质干细胞增殖能力的影响存在浓度相关性，25 mg/L为最佳标记浓度。磁共振成像T2*WI序列更适合追踪菲立磁标记的人骨髓间质干细胞。

关键词: 菲立磁, 标记, 骨髓间质干细胞, 磁共振成像, 增殖

Abstract:

BACKGROUND: Conventional detection methods of stem cell tracking are lack of succession and noninvasion, because most of them need animal organizations to be inspected.
OBJECTIVE: To identify the effect of Feridex on human bone marrow mesenchymal stem cells (BMSCs) activities and determine suitable concentration of Feridex and screen sequence of magnetic resonance imaging (MRI) scan, through labeling BMSCs with Feridex ex vivo.
METHODS: Passage 3 BMSCs in good condition were labeled with Feridex of different concentrations (100, 50, 25, 12.5 mg/L). The culture was performed for 24-48 hours. Blank control group included Passage 3 BMSCs that were not labeled. Proliferation of BMSCs of the two groups was detected with methyl thiazolyl tetrazolium (MTT), and growth curve was made. The labeling was identified with Prussian blue staining. MRI scan of BMSCs labeled with Feridex was performed to determine the suitable sequence of MRI scan.
RESULTS AND CONCLUSION: There was no effect of Feridex with concentration ≤25 mg/L on BMSCs, and the result of labeling with concentration of 25 mg/L was optimal. Feridex with concentration≥ 50 mg/L would obviously inhibit the proliferation of BMSCs. Results of MRI scan revealed that MRI signal of BMSCs Feridex labeled for 24 and 48 hours or the filial Passage degraded in all sequence T1WI, T2WI and T2*WI3, the change of T2*WI was mostly obvious. These results indicate that Feridex can be used for ex vivo labeling of BMSCs for succession research. The effect of Feridex on BMSCs was in concentration dependablity, and concentration of 25 mg/L was optimal. The result of sequence T2*WI of MRI scan was more suitable to trace Feridex-labeled human BMSCs

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