Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (45): 8413-8418.doi: 10.3969/j.issn.2095-4344.2012.45.010

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Transfection of an eukaryotic expression vector expressing brain derived neurotrophic factor into human umbilical cord mesenchymal stem cells

Sun Zhao-ming1, Yuan Yuan2, Pan Shu-mao2, Zhang Ju-hua2, Wang Yuan-li3, Tang Guo-tai2, Guan Mao-wu2, Chen Hong-guang2   

  1. 1Department of Neurology, 2Department of Neurosurgery, 3Department of Obstetrics and Gynecology, Yuhuangding Hospital of Yantai, Yantai 264000, Shandong Province, China
  • Received:2012-06-01 Revised:2012-08-31 Online:2012-11-04 Published:2012-11-04
  • Contact: Yuan Yuan, Doctor, Department of Neurosurgery, Yuhuangding Hospital of Yantai, Yantai 264000, Shandong Province, China yuan_xing_yuan@yahoo.com.cn
  • About author:Sun Zhao-ming☆, Doctor, Attending physician, Department of Neurology, Yuhuangding Hospital of Yantai, Yantai 264000, Shandong Province, China sunzhaoming@tom.com

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Abstract:

BACKGROUND: Stem cells transplantation is the research hot spot in the area of nerve injury repair. At present, most researches have focused on neural stem cells and bone marrow stem cells transplantation, and there are rare reports on the research of umbilical cord mesenchymal stem cells, but the umbilical cord mesenchymal stem cells have the advantages of rich source and little ethical restrictions.
OBJECTIVE: To construct an eukaryotic expression vector expressing brain derived neurotrophic factor, and to transfect it into the umbilical cord mesenchymal stem cells and establish a cell model of brain derived neurotrophic factor.
METHODS: Firstly, the c-DNA of brain derived neurotrophic factor was gained by reverse transcription-PCR and then inserted into multi-clone site of pcDNAⅢ vector to construct the expression vector of brain derived neurotrophic factor pcDNAⅢ. Then the vector was used to transtect umbilical cord mesenchymal stem cells. At last, the expression level of brain derived neurotrophic factor in the umbilical cord mesenchymal stem cells was tested by Western blot; the cultured PC12 cells and the embryo rat pallium neurons were added into the umbilical cord mesenchymal stem cells containing brain derived neurotrophic factor to test the biological activity of brain derived neurotrophic factor.
RESULTS AND CONCLUSION: The eukaryotic expression vector expressing brain derived neurotrophic factor was successfully constructed, and this vector could highly express the brain derived neurotrophic factor protein in umbilical cord mesenchymal stem cells. In vitro testing showed the brain derived neurotrophic factor protein had the biological activity. It indicates that the umbilical cord mesenchymal stem cells can be used as the eukaryotic expression vector of brain derived neurotrophic factor for research on the nerve injury repair.

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