Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (37): 6847-6851.doi: 10.3969/j.issn.1673-8225.2010.37.002

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Construction of recombinant adenovirus vector containing mechano-growth factor gene and its expression in rat osteoblasts

Qiu Min, Tang Li-ling   

  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College of Chongqing University, Chongqing  400044, China
  • Online:2010-09-10 Published:2010-09-10
  • Contact: Tang Li-ling, Doctor, Professor, Doctoral supervisor, Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College of Chongqing University, Chongqing 400044, China lilingtang@yahoo.com.cn
  • About author:Qiu Min★, Studying for master’s degree, Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College of Chongqing University, Chongqing 400044, China
  • Supported by:

    the General Program of National Natural Science Foundation of China, No. 30600130*, 30970701*; Science and Technology Innovation Foundation for Post-graduate in Chongqing University, No. 200904AIA0010308*

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Abstract:

BACKGROUND: Studies have confirmed that mechano-growth factor (MGF) can activate satellite cell and facilitate myoblast proliferation, which play an important role in treating muscle loss, preventing myocardial damage and repairing nerve injury. Mechanical stretch can accelerate MGF expression in osteoblast, however, the influence mechanisms of MGF on physiological and biochemical behavior of bone tissues remain poorly understood. 
OBJECTIVE: Osteoblasts were transfected with recombinant adenovirus vector containing MGF gene to observe the expression of MGF in osteoblasts.
METHODS: The MGF coding sequence was cloned to the pAdtrack -CMVplasmid to construct pAdtrack-MGF. Then pAdtrack-MGF was transformed into E. Coli BJ5183 carrying backbone plasmid already, and in which following co-transformation with the backbone vector pAdEasy-1. The homologous recombinant was transfected into human embryo kidney cells 293A through the lipofectamine to pack the adenovirus. The identified recombinant adenovirus (Ad/MGF) was amplified in 293A cells. The infection efficiency was measured by fluorescent tracer technique, and the results of recombinant adenovirus vector infection were identified by RT-PCR.
RESULTS AND CONCLUSION: The title of recombinant adenovirus was 8.5×108 pfu/ mL. Osteoblasts infected by Ad/MGF could over express MGF. When multiplicity of infection was equal to 100, the vector had the best efficacy of infection. The recombinant adenovirus vector containing MGF gene could transfect the rat osteoblasts successfully.

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