Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells
    8.
    发明授权
    Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells 失效
    生产高滴度病毒和高效逆转录病毒介导的哺乳动物细胞转导的方法

    公开(公告)号:US06506604B2

    公开(公告)日:2003-01-14

    申请号:US09944411

    申请日:2001-09-04

    IPC分类号: C12N15867

    摘要: 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 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.

    摘要翻译: 本发明提供了一种新型的逆转录病毒包装系统,其中在哺乳动物细胞中稳定或瞬时转染之后,由高表达质粒产生逆转录病毒包装质粒和可包装载体转录物。 在这些转染的哺乳动物细胞中产生高滴度的重组逆转录病毒,然后可以通过共培养或上清液感染来转导哺乳动物靶细胞。 本发明的方法包括使用新的逆转录病毒包装质粒和载体以高效率通过共培养或上清液感染来转导包含T细胞和人造血干细胞的原代人细胞与外来基因。 本发明可用于快速生产高滴度病毒上清液,并且可以通过常规方法转导难以转导的高效细胞。