SUSTAINED AND CONTINUOUS PRODUCTION OF HIGH TITERS OF RECOMBINANT VIRAL VECTORS AND TRANSDUCED TARGET CELLS FOR USE IN GENE THERAPY

摘要

This invention provides improved methods for producing high titers of recombinant viral vectors by culturing producer cells in a hollow fiber bioreactor. The titer of virus in the extra fiber space (EFS) of the hollow fiber bioreactor is sufficient to transduce target cells at a multiplicity of infection sufficient to render the transduced target cells useful for genetic therapy. In a preferred embodiment, target cells are transduced with the EFS medium from a hollow fiber bioreactor in which a producer cell line, which releases packaged retroviral vectors into the EFS, has been cultured. Lymphocytes derived from an adenosine deaminase (ADA)-deficient individual that have been transduced with EFS medium that contains retroviral vectors that include RNA that encodes ADA, express ADA at a rate comparable to that of cells from an individual who does not have ADA deficiency.