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China Journal of Orthopaedics and Traumatology ; (12): 1023-1027, 2013.
Article Dans Chinois | WPRIM | ID: wpr-250705

Résumé

<p><b>OBJECTIVE</b>To explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.</p><p><b>METHODS</b>Six clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.</p><p><b>RESULTS</b>1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).</p><p><b>CONCLUSION</b>There is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.</p>


Sujets)
Animaux , Femelle , Humains , Mâle , Rats , Protéines adaptatrices de la transduction du signal , Génétique , Métabolisme , Prolifération cellulaire , Cellules cultivées , Protéines du cytosquelette , Génétique , Métabolisme , Thérapie génétique , Vecteurs génétiques , Génétique , Métabolisme , Protéines à domaine LIM , Génétique , Métabolisme , Lentivirus , Génétique , Métabolisme , Cellules souches mésenchymateuses , Biologie cellulaire , Métabolisme , Virologie , Ostéoporose , Génétique , Thérapeutique , Rat Sprague-Dawley
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