发明名称 |
Efficient gene transfer into primary lymphocytes |
摘要 |
The present invention pertains to a method for efficiently introducing exogenous genes into primary lymphoid cells without drug selection which comprises the steps (a) deriving a retroviral vector and a helper cell combination that will yield a level of virus production in the range from 5x106 to 5x107 units/ml by transfecting a vector into a helper cell followed by selection, isolation of cell clones, and determination of viral titers to identify which virus-producing cell lines produce a virus titer in the range from 5x106 to 5x107 units/ml; (b) isolating a lymphoid cell subpopulation which can repopulate a specific lymphoid lineage or is a long-lived population by treating a suspension of lymphoid cells with a monoclonal antibody which removes undesired lymphoid cells to obtain an enriched lymphoid subpopulation; (c) culturing the enriched lymphoid subpopulation from step (b) with growth factors specific to the lymphoid subpopulation; (d) co-cultivating the lymphoid subpopulation from step (c) with a lawn of irradiated virus-producing cell line from step (a) to produce an infected lymphoid subpopulation; and (e) harvesting the infected lymphoid subpopulation. |
申请公布号 |
AU7477394(A) |
申请公布日期 |
1995.03.27 |
申请号 |
AU19940074773 |
申请日期 |
1994.08.01 |
申请人 |
UNIVERSITY OF MEDICINE & DENTISTRY OF NEW JERSEY |
发明人 |
JOSEPH DOUGHERTY;MING-LING KUO;NATALIE SUTKOWSKI;RON YACOV |
分类号 |
C12N15/09;C12N5/10;C12N15/85;C12N15/867;C12N15/87 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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