Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6218187

Title:Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells
Inventors:Finer, Mitchell H.; San Carlos, CA, USA
Dull, Thomas J.; San Francisco, CA, USA
Zsebo, Krisztina M.; Woodside, CA, USA
Cooke, Keegan; Palo Alto, CA, USA
Farson, Deborah A.; Oakland, CA, USA
Summary:This invention introduces a high efficiency retroviral packaging system which may be used to transduce mammalian hematopoietic stem cells with retroviral supernatants and T or B cells with foreign genes. Described herein is a novel retroviral packaging system produced by transient transfection in which retroviral packaging plasmids and packagable vector transcripts are produced from high expression plasmids after stable or transient transfection in mammalian cells. Claims of the invention include high titers of recombinant retrovirus which are produced in transfected mammalian cells and which then transduce a mammalian target cell by cocultivation or supernatant infection. Therapeutic applications include the rapid production of high titer viral supernatants, and the efficient transduction of cells that are refractory to transduction by conventional means.
Abstract:The invention provides a novel retroviral packaging system, in which retroviral packaging plasmids and packagable vector transcripts are produced from high expression plasmids after stable or transient transfection in mammalian cells. High titers of recombinant retrovirus are produced in these transfected mammalian cells and can then transduce a mammalian target cell by cocultivation or supernatant infection. The methods of the invention include the use of the novel retroviral packaging plasmids and vectors to transduce primary human cells, including T cells and human hematopoietic stem cells, with foreign genes by cocultivation or supernatant infection at high efficiencies. The invention is useful for the rapid production of high titer viral supernatants, and to transduce with high efficiency cells that are refractory to transduction by conventional means.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6218187
Application Number:US1999000266596
Date Filed:11/03/1999
Date Published:17/04/2001
Assignee:Cell Genesys, Inc., Foster City, CA, USA


 
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