MAMMALIAN CELL TRANSDUCTION FOR USE IN GENE THERAPY

摘要

The present invention relates to a method for the ex vivo transduction of mammalian cells, in particular to the transduction of bone marrow stromal cells. These cells can be transduced with a gene of interest, in particular a B-domain deleted human factor VIII gene. In the latter case, the transduced cells can be used to treat hemophilia A. The method for the ex vivo transduction of bone marrow stromal cells with the human factor VIII gene comprises provision of an intron-based retroviral vector comprising a B-domain deleted human factor VIII cDNA (designated as MFG-FVIII DELTA B); pseudotyping the said vector with the Gibbon ape leukemia virus (GALV) envelope; transducing bone marrow stromal cells with the said pseudotyped vector by pre-incubating the cells for a suitable period of time in cell culture medium without phosphate and subsequently adding a vector-containing supernatant, optionally supplemented with transduction additives to the cells, followed by centrifuging the mixture thus obtained; and optionally repeating the two previous steps.