Differentiation of rabbit bone marrow mesenchyme stem cells co-transfected by TGF-β3 and BMP-7 adenoviral vectors into nucleus pulposus-like cells

English Keywords:BMSCs  Adenovirus  TGF-β3  BMP-7  Seed cell

English Abstract:

  【Abstract】 Objectives: To induce rabbit bone marrow mesenchyme stem cells(BMSCs) differentiating into nucleus pulposus-like cells(NPCs) by co-transfecting with TGF-β3 and BMP-7 adenoviral vectors. Methods: BMSCs were divided into five groups: A, blank control group; B, transfected by blank GFP adenoviral vector group; C, transfected by TGF-β3 adenoviral vector group; D, transfected by BMP-7 adenoviral vector group; E, co-transfected by TGF-β3 and BMP-7 adenoviral vectors group. The transfection result was observed by fluorescence microscope. After 14 days′ cultivation, the expressions of TGF-β3 and BMP-7 protein(group A, C, D, E) and ACAN, Collagen Ⅰ, Collagen Ⅱ, Collagen Ⅹ and SOX9(all group) in mRNA level were measured by Western blot and Realtime PCR respcetively. Results: 14 days after culture of co-transfected BMSCs, the shape of most cells changed obviously. Western blot showed higher expression of TGF-β3 and BMP-7 proteins in group E. The expressions of ACAN, Collagen Ⅰ, Collagen Ⅱ and SOX9 gene were much higher than those control group(group A, B)(P<0.05), and the expression of Collagen Ⅱ in co-transfected group was higher compared with single-transfected groups(P<0.05). The expression of Collagen Ⅹ in TGF-β3 transfected group and co-transfected group was obviously lower than that of BMP-7 transfected group and control group(P<0.05). Conclusions: The rabbit BMSCs are able to express TGF-β3 and BMP-7 proteins and differentiate into NPCs after co-transfected by TGF-β3 and BMP-7 adenoviral vectors.