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    Bone Marrow Derived Macrophage (BMDM) differentiation and maintenance

    Narayana Yadavalli, Shawn M. Ferguson

    Published: 2023-08-05 DOI: 10.17504/protocols.io.j8nlkw7bwl5r/v1
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    Abstract

    This protocol describes the isolation of bone marrow derived macrophages.

    Attachments

    724-1815.docx

    Steps

    1.

    Note
    Note : For mouse bone marrow-derived macrophage (BMDMs) primary cultures, each experiment involved age and sex matched C57BL/6 mice between 3-9 months of age. Lrrk2 KO (Lrrk2tm1.1Mjff) and G2019S (Lrrk2tm1.1Hlme) homozygous knockin mice were obtained from The Jackson Laboratory76.
    Euthanize mice via CO2 or isoflurane inhalation and cervical dis.

    2.

    Collect femurs and flush cavities with 5mL ice-cold PBS.

    3.

    Then, collect bone marrow cells by centrifugation 0.3rcf.

    4.

    Resuspend the resulting pellet and differentiate for 6 days in culture media containing: DMEMF12 supplemented with 20% FBS, 20% L929 conditioned media, 1% penicillin-streptomycin and 1% GlutaMAXTM.

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