Rabbit Bone Marrow Mesenchymal Stem Cells Expanded In Vitro And Experiments Of BMP-2 Eukaryotic Expression Plasmid Transfection

Objective:(1) Transfection of bone marrow culture needed to mesenchymal stem cells (bone marrow mesenchymal stem cells, BMSCs), (2)Through the use of cationic liposomes pcDNA3.1 (-)-mediated transfection method to BMSCs, BMSCs were stably transfected cells. (3) The use of immunohistochemistry, WesternBlot pcDNA3.1-hBMP2 detected in rabbit BMSCs in protein expression.Methods:(1) Adherent BMSCs cultured and amplified required from the morphology, surface markers by flow cytometry. (2) We have been successfully constructed recombinant plasmid pcDNA3.1-hBMP2 target cells in vitro transfection and G418 selection using stable cell. (3) By immunohistochemistry, WesternBlot assay of protein expression in BMSCs.Result:(1) After cultivation, expansion of bone marrow mesenchymal stem cells, P2, P3 can be used as hBMP2 transfected target cells. (2) Cationic lipid transfection pcDNA3.1-hBMP2 of MSCs by G418 screening by immunohistochemistry, WesternBlot assay, transfection pcDNA3.1-hBMP2 of rabbit bone marrow mesenchymal stem cells have a large number of hBMP2 mRNA transcription and related proteins, without transfection of bone marrow mesenchymal stem cells no expression.Conclusion:(1) Rabbit bone marrow mesenchymal stem cells can be used as hBMP2 transfected target cells. (2) The eukaryotic hBMP2 plasmid in rabbit bone marrow mesenchymal stem cells are related to expression. (3) This study further BMP2 gene transfected MSCs proliferation and stable expression in vitro and in animal studies to accelerate new bone formation in distraction osteogenesis in experimental basis.