Human hepatocyte growth factor gene modified bone marrow mesenchymal stem cells in rats.docVIP

 PAGE \* MERGEFORMAT 14 Human hepatocyte growth factor gene modified bone marrow mesenchymal stem cells in rats Of: Zhu Yuerong, Xingji Cheng, Han Ping, Qiu [Abstract] Objective: To construct containing human hepatocyte growth factor (hHGF) gene in mesenchymal stem cells (MSCs), to obtain high expression hHGF of MSCs. METHODS: containing hHGF adenovirus vector pDC316 HGF IRES EGFP. The same source obtained with recombinant adenovirus hHGF Ad HGF. to express only GFP recombinant adenovirus Ad GFP as a control, in vitro respectively, Ad GFP, Ad HGF infected MSCs. fluorescence activated cell sorting detection of infected cells, genes expression efficiency. enzyme-linked immunosorbent assay detection of transduced cells the expression of the culture supernatant hHGF concentration and duration. Results: The adenoviral expression vector by restriction digestion after the hHGF 2.2 kb and 5.2 kb gene fragment The linearized vector fragment with the gene fragment sequencing results hHGF cDNA library sequence. viral titer of purified liquid Ad HGF can reach 8.02 1010 pfu / ml.FACS detect HGF / MSCs of GFP-positive rate of 95.19%. HGF / MSCs in cell culture medium hHGF can continue to express at least 14 d, the highest concentration of 99 ng / ml. Conclusion: The expression of adenovirus vector in the correct, high titer recombinant adenovirus Ad HGF solution, after transduction MSCs to get efficient, stable and sustained expression of hHGF. The vector to test the design requirements for further in vivo studies provide an experimental basis. [Keywords:] mesenchymal stem cell; liver cell growth factor, adenovirus vector [Abstract] Objective: To construct the adenoviral expression vector system containing human HGF cDNA, and produce the virus containing hHGF, and to further study the transducing efficiency and the expression of HGF in MSCs. Methods: The HGF cDNA was amplificated from the expression plasmid pCMV HGF by PCR, and subcloned into the same site of th