中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (19): 3520-3524.doi: 10.3969/j.issn.1673-8225.2012.19.021

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

 氯甲基苯甲酰胺标记大鼠肾脂肪囊来源脂肪间充质干细胞的可行性*★

訾利影1，郑红光2，张德伟2，梅煜明2   

  1. 1辽宁医学院沈阳军区总医院研究生培养基地，辽宁省锦州市121011；2解放军沈阳军区总医院，辽宁省沈阳市110016
  • 收稿日期:2012-01-28 修回日期:2012-02-24 出版日期:2012-05-06 发布日期:2012-05-06
  • 通讯作者: 郑红光，主任医师，教授，硕士生导师，解放军沈阳军区总医院肾脏病科，辽宁省沈阳市 110016
  • 作者简介:訾利影★，女，1984年生，河北省廊坊市人，汉族，辽宁医学院在读硕士，主要从事肾脂肪囊来源脂肪间充质干细胞的示踪及归巢研究。ziliyingyinglizi@163.com
  • 基金资助:

    2011年辽宁省攻关课题。

Feasibility of chlormethyl-benzamidodialkylcarbocyanine labeling rat kidney fat capsule adipose-derived mesenchymal stem cells 

Zi Li-ying1, Zheng Hong-guang2, Zhang De-wei2, Mei Yu-ming2   

  1. 1Postgraduate Culture Base, the General Hospital of Shenyang Military Region, Liaoning Medical University, Jinzhou 121011, Liaoning Province, China; 2the General Hospital of Shenyang Military Region, Shenyang  110016, Liaoning Province, China
  • Received:2012-01-28 Revised:2012-02-24 Online:2012-05-06 Published:2012-05-06
  • Contact: Zheng Hong-guang, Chief physician, Professor, Master’s supervisor, the General Hospital of Shenyang Military Region, Shenyang 110016, Liaoning Province, China
  • About author:Zi Li-ying★, Studying for master’s degree, Postgraduate Culture Base, the General Hospital of Shenyang Military Region, Liaoning Medical University, Jinzhou 121011, Liaoning Province, China ziliyingyinglizi@163.com
  • Supported by:

    Key Technologies Research and Development Program of Liaoning Province in 2011*

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摘要：

背景：氯甲基苯甲酰胺被认为是目前较为稳定的细胞荧光标记物质。
目的：观察荧光染料氯甲基苯甲酰胺标记肾脂肪囊来源脂肪间充质干细胞的标记效果及其对细胞形态、生长及增殖能力等生物学特性的影响。
方法：采用胶原酶消化法分离、培养SD大鼠肾脂肪囊来源脂肪间充质干细胞，以氯甲基苯甲酰胺细胞标记液标记第3代肾脂肪囊来源脂肪间充质干细胞。
结果与结论：荧光显微镜下观察氯甲基苯甲酰胺标记的肾脂肪囊来源脂肪间充质干细胞呈红色荧光，细胞标记率达95%以上，标记前后细胞形态、生长及增殖等方面无明显变化。经多次传代培养后，红色荧光细胞数量及强度有所下降，但细胞传代至第8代，氯甲基苯甲酰胺标记后的第15天细胞标记率仍可达50%以上。表明氯甲基苯甲酰胺可有效标记肾脂肪囊来源脂肪间充质干细胞，且方法简单，标记率高，对细胞无毒性，不影响细胞生物学特性。
 

关键词: 氯甲基苯甲酰胺(CM-DiI), 肾脂肪囊, 脂肪间充质干细胞, 标记, 大鼠

Abstract:

BACKGROUND: Chlormethyl-benzamidodialkylcarbocyanine (CM-DiI) has been considered a relatively stable lipophilic fluorescent dye.
OBJECTIVE: To investigate the labeling effects of CM-Dil for kidney fat capsule adipose-derived mesenchymal stem cells and its effects on cell morphology, growth and proliferation.
METHODS: Sprague-Dawley rat kidney fat capsule adipose-derived mesenchymal stem cells were isolated by collagenase digestion. Passage 3 kidney fat capsule adipose-derived mesenchymal stem cells were labeled by CM-Dil.
RESULTS AND RESULTS: Under fluorescent microscope, CM-Dil-labeled kidney fat capsule adipose-derived mesenchymal stem cells exhibited red fluorescence, with a labeling rate of over 95%. There were no obvious changes in cell morphology, growth and proliferation before and after labeling. After several passages, the number and staining intensity of cells with red fluorescence were decreased. At 15 days after CM-Dil labeling, over 50% of passaged cells were labeled. These suggest that CM-Dil can effectively label fat capsule adipose-derived mesenchymal stem cells and this method is easy-to-operate, with a high labeling rate, produces no cytotoxicitry and does not influence cell biological characteristics. 

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