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  • Title: Long-term cultures of mouse bone marrow cells: a model for studying the generation of natural killer cells.
    Author: Ayroldi E, Cannarile L, D'Adamio F, Delfino D, Migliorati G, Riccardi C.
    Journal: Nat Immun; 1992; 11(6):317-27. PubMed ID: 1477494.
    Abstract:
    We investigated the generation of natural killer (NK) cells, using a long-term bone marrow culture (LTBMC) system. Mouse bone marrow cells were cultured for 2 weeks in complete medium without growth factors to obtain an enriched population of NK precursor cells. When these cells were recultured in the presence of interleukin-2 (IL-2) and conditioned medium (CM) from LTBMC, lytic NK cells were generated within 4 days. Replacing CM with fresh medium, before adding IL-2, decreased NK cell generation markedly, suggesting that endogenous factors present in CM were necessary for IL-2 induction of NK cells. NK cell precursors were also cultured with a combination of IL-2 and interferon-gamma (IFN-gamma) or IL-2 and tumor necrosis factor-alpha (TNF-alpha), but no CM. Results show that IFN-gamma and TNF-alpha were able to substitute CM. The addition of anti-IFN-gamma or anti-TNF-alpha antibodies to LTBMC cells, cultured in the presence of IL-2 and CM, inhibited cytotoxicity induction in a dose-dependent manner. The data indicate that IFN-gamma and TNF-alpha production may be required for IL-2 induction of NK activity, and are consistent with the hypothesis that NK generation involves collaboration between IL-2 and other bone marrow microenvironmental growth factors.
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