Combination Gene Transfection Of TGF-β3and BMP-2Mediated By Lentivirus Induce Chondrogenicdifferentiation Of Rabbit Bone Marrow Mesenchymal Stem Cells

Objective Combination gene transfection of TGF-β3/BMP-2mediated by recombinant lentivirus induce chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells(BMSCs), and compared with TGF-β3single gene transfecting.Methods BMSCs were isolated and cultured using a whole bone marrow adherent method, then detected cells’surface antigen by Flow cytometry, respectively transferred negative control (N group), TGF-β3single gene (T group), TGF-(33/BMP-2combination gene (TB group), observed cell morphology of each group, the expression of SOX-9, type Ⅱ collagen (COL2), aggrecan(ACAN) by RT-PCR and Western Blot respectively in gene and protein level, moreover, toluidine blue staining and type II collagen immunocytochemistry were performed to detect the extracellular matrix.Results The BMSCs revealed uniformly negative for the early hematopoietic stem cell marker(CD34,2.07%) and the leucocytes surface antigen (HLA-DR,1.73%), positively for the hyaluronan receptor(CD44,82.79%). The differentiated cells took a shape of polygonal, square or disc-like, the expression of chondrogenic related markers such as sox-9, COL2and AC AN were upregulated in T group and TB group than in C group and N group (P<0.05), furthermore, those markers were synthetized significantly more in TB group than in T group (P<0.01).Conclusion These findings indicated that BMSCs differentiated into chondrocytes induced by TGF-β3/BMP-2combination gene, this method was superior to TGF-β3single gene transfection.