Study on Retrovirus-Mediated Murine GM-CSF cDNA Transfer and Expression in Bone Marrow Stromal Cells

In order to investigate the feasibility of retrovirus-mediated GM-CSF gene transfer into bone marrow stromal cells (BMSC), granulocyte/macrophage colony-stimulating factor (GM-CSF) cDNA was recombined with retroviral vector pLXSN. After this recombinant plasmid was transfected into retro-virus packaging cell line PA317 by eletroporation,. G418 resistant clones could produce GM-CSF cDNA recombinant retrovirus successfully which could infect stromal target cells and make them form typical resistant clones in G418 selective medium. In the genome of these infected target cells, NeoR gene and GM-CSF cDNA were successfully integrated, the GM-CSF mRNA expression in the BMSC was detected by in situ hybridization and the specific activity of GM-CSF in the cultural medium was assessed. These results demonstrated that BMSC transfected with GM-CSF cDNA could significantly augment GM-CSF in vitro and might become target cells of gene therapy, and provided a basis for further investigation on hematopoietic reconstitution of BMSC mediated GM-CSF gene therapy.