中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6651-6656.doi: 10.3969/j.issn.1673-8225.2010.36.001

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

mdx小鼠与C57小鼠骨髓基质细胞体外分化能力的比较

冷  雁，张为西，周  琛，郑振扬，张  成   

  1. 中山大学附属第一医院神经科，广东省广州市   510080
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 通讯作者: 张为西，博士，教授，硕士生导师，中山大学附属第一医院神经科，广东省广州市 510080 weixizhang@qq.com
  • 作者简介:冷雁★，女，1984年生，湖南省永州市人，汉族，中山大学附属第一医院在读硕士，主要从事骨髓干细胞与神经肌肉疾病的研究。 Lengyan19840117@sina.com
  • 基金资助:

    课题由2项国家自然基金资助：GILZ对骨髓间质干细胞分化的调控及其机制的研究(30870852)；GILZ转导的MSC移植治疗DMD模型的研究(30971026)。

Bone marrow stromal cells differentiation potentials in vitro from mdx mice versus C57 mice

Leng Yan, Zhang Wei-xi, Zhou Chen, Zheng Zhen-yang, Zhang Cheng   

  1. Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou  510080, Guangdong Province, China
  • Online:2010-09-03 Published:2010-09-03
  • Contact: Zhang Wei-xi, Doctor, Professor, Master’s supervisor, Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China weixizhang@qq.com.
  • About author:Leng Yan★, Studying for master’s degree, Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China Lengyan19840117@sina.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30971026*，30870852*

PDF

320

被引次数

7

摘要：

背景：肌营养不良症是一种渐进性致死性X连锁隐性遗传性肌肉疾病，目前无特效治疗。肌营养不良症模型鼠(mdx小鼠)的骨髓基质细胞增殖及定向分化能力是否正常，自身骨髓移植是否合适还有待研究。
目的：观察mdx小鼠骨髓基质细胞体外培养时的增殖及多向分化能力。
方法：取mdx小鼠与C57小鼠胫股骨骨髓基质细胞体外培养，经吉姆萨染色后观察其形成成纤维细胞集落形成单位的能力；通过不同诱导液使骨髓基质细胞定向分化为成骨、成脂、成肌细胞，观察其形态学特性；并分别用Von kossa 染色、油红O染色、免疫荧光检测desmin阳性细胞对已分化细胞进行鉴定和分化率比较；培养1周时，提取分化细胞总RNA，反转录后，用real-time PCR检测各分化细胞相关基因表达。
结果与结论：mdx小鼠骨髓基质细胞形成的成纤维细胞集落形成单位数目和体积均小于C57小鼠。其成骨、成肌分化的效率均明显低于C57小鼠(P < 0.01)，两种小鼠的骨髓基质细胞成脂分化效率差异无显著性(P > 0.05)。real-time PCR检测结果显示，与C57小鼠相比，mdx小鼠的骨髓基质细胞成骨、成肌基因表达均有不同程度下降，而成脂基因表达无明显差异。结果提示，mdx小鼠的骨髓基质细胞体外培养时的增殖及定向分化能力较C57小鼠下降，与Dystrophin 基因缺失有关，mdx小鼠自体骨髓移植将会受限。

关键词: 小鼠, 骨髓基质细胞, 诱导, 分化, 肌营养不良症

Abstract:

BACKGROUND: Duchenne’s muscular dystrophy (DMD) is a fatal recessive X-linked form of muscular dystrophy characterized by progressive muscular degeneration with no certain treatment. However, the proliferation and multipotent differentiation potential of bone marrow stromal cells (BMSCs) from DMD model mice (mdx mice) and the effectiveness of BMSCs self-transplantation are needed to be further studied.
OBJECTIVE: To investigate the proliferation and multipotent differentiation potential of BMSCs from mdx mice during in vitro culture.
METHODS: Both C57 and mdx mice were killed to obtain the BMSCs for culturing in vitro. After Giemsa’s staining, the colony forming unit-fibroblast (CFU-f) of BMSCs assay was performed. With specific inductive mediums, we succeeded to induce the BMSCs to differentiate into osteogenesis, adipogenesis, myogenesis respectively. Their morphological characteristics were observed with microscope. Von kossa staining, oil red O staining and immunofluorescence for desmin were utilized to identify the differentiated BMSCs respectively and quantify their differentiation efficiency. After the BMSCs were induced for 1 week, the cellular total RNA of differentiated BMSCs was extracted, and then reverse transcription was performed. Real-time polymerase chain reaction (PCR) was used to quantify the gene expression of differentiated BMSCs.
RESULTS AND CONCLUSION: Compared with C57 mice, both number and volume of CFU-f of BMSCs from mdx mice were smaller. Compared to C57 mice, the efficiency of osteogenesis and myogenesis of BMSCs from mdx mice was significantly lower (P < 0.01). However, the efficiency of adipogenesis of BMSCs from both groups had no statistic difference (P > 0.05). Real-time PCR showed that both of osteogenic and myogenic gene expression of BMSCs from mdx mice decreased respectively compared to C57 mice. However, adipogenic gene expression from two groups had no difference. The results indicated that proliferation and multipotent differentiation potentials of BMSCs from mdx mice declined compared with C57 mice. The defection of Dystrophin gene may contribute to it. Therefore, the autoplastic BMSCs transplantation of mdx mice will be interfered.

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