Retrovirus Mediated GM-CSF Gene Transfer into NIH3T3 Cells

Recombinant retrovirus containing bacterial neomycin-resistant (Neo R) gene and granulocyte-macrophage colony-stimulating factor was transfered into retrovirus-packaging cell PA317 by electropo-ration. The supernatant of the transfected cells was used to infect NIH3T3 cells. Transfection efficiency was determined by in situ PCR detection of Neo R gene and colony formation by G418 resistant cells. Southern blot analysis showed integration of GM-CSF cDNA into host chromosome. In situ hybridization showed GM-CSF mRNA expression and biological assay showed GM-CSF activity in the culture medium.