abstract |
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. |