Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (23): 4303-4306.doi: 10.3969/j.issn.1673-8225.2011.23.028

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Adenovirus carrying adrenomedullin gene for transfection of bone marrow mesenchymal stem cells in vitro and their protein expression

Lin Mao-hai1, Su Jin-zi2, Cai Wen-qin2, Zheng Li-yu2, Xu Chang-sheng3, Wang Hua-jun3   

  1. 1Department of Cardiology, The First Hospital of Putian City, Putian  351100, Fujian Province, China
    2Department of Cardiology, The First Affiliated Hospital, Fujian Medical University, Fuzhou  350005, Fujian Province, China
    3Fujian Institute of Hypertension Research, The First Affiliated Hospital of Fujian Medical University, Fuzhou  350005, Fujian Province, China
  • Received:2010-12-19 Revised:2011-05-07 Online:2011-06-04 Published:2011-06-04
  • About author:Lin Mao-hai★, Master, Attending physician, Department of Cardiology, The First Hospital of Putian City, Putian 351100, Fujian Province, China lindrogon@hotmail.com
  • Supported by:

    the Scientific Research Foundation of First Affiliated Hospital of Fujian Medical University for Leaders in the Field of Learning, No.JXK200704*

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Abstract:

BACKGROUND: Adrenomedullin is one of potential growth factor that can promote cellular differentiation, growth and reduce apoptosis.
OBJECTIVE: To investigate the possibility of bone marrow mesenchymal stem cells transfected by adenovirus vector carrying adrenomedullin (Ad-ADM), and to determine the protein expression of adrenomedullin following transfection.
METHODS: The 293A cells were cultivated and the Ad-ADM was amplified. The experiment was divided into three groups: control, blank vector and Ad-ADM transfection. Cells in three groups were cultured with serum-free L-DMEM, Ad-lacZ and Ad-ADM, respectively after they reached 50%-60% confluence. The virus titer of amplified Ad-ADM was detected by plaque methods. Bone marrow mesenchymal stem cells were isolated and expanded using the preplating methods. The efficiency of Ad-GFP transfected mesenchymal stem cells in vitro was determined. Adrenomedullin mRNA expression was detected with reverse transcription-polymerase chain reaction following transfection.
RESULTS AND CONCLUSION: The virus titer of amplified Ad-ADM was 1.1×109 pfu/mL by plaque methods. After bone marrow mesenchymal stem cells were infected with Ad-ADM, the expression of adrenomedullin mRNA was significantly increased. The expression of adrenomedullin mRNA was not observed in the control group and empty vector group. Quantitative results showed that the expression gradually increased from day 1, reached the peak at day 7, and was still detectable at day 11. The Ad-ADM can successfully transfect bone marrow mesenchymal stem cells. Following transfection, bone marrow mesenchymal stem cells highly express adrenomedullin.

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