Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (36): 6651-6656.doi: 10.3969/j.issn.1673-8225.2010.36.001

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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*

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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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