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Bone marrow cell culturing in double diffusion chambers

P Pfeffer, A Boyum

    Scandinavian Journal of Haematology
    |February 1, 1977
    PubMed
    Summary
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    A novel double diffusion chamber technique (DDC) was developed to study bone marrow cells (BMC). Spleen cells stimulated BMC growth, while other BMC inhibited it, revealing complex cellular interactions.

    Area of Science:

    • Hematology
    • Cell Biology
    • Immunology

    Background:

    • Bone marrow cells (BMC) are crucial for hematopoiesis.
    • Understanding cellular interactions influencing BMC proliferation is vital.
    • Existing methods may not fully capture regulatory microenvironments.

    Purpose of the Study:

    • To establish and validate a double diffusion chamber (DDC) technique.
    • To investigate the influence of diffusible factors from spleen cells and BMC on bone marrow cell growth.
    • To analyze the effects on colony-forming units (CFU-S, CFU-C), granulocytes, and macrophages.

    Main Methods:

    • A DDC system was created with two compartments separated by a Millipore membrane.
    • Mouse bone marrow cells were cultured in one compartment.

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  • The other compartment contained medium (control), spleen cells, or BMC as regulators.

    • Main Results:

    • BMC proliferated rapidly in control DDC, with only a 20% reduction in yield compared to single chambers.
    • Spleen cells significantly stimulated the growth of CFU-S and CFU-C.
    • BMC in the regulator compartment suppressed granulopoiesis in the target compartment.

    Conclusions:

    • The DDC technique effectively supports BMC culture and allows investigation of cellular interactions.
    • Diffusible factors from spleen cells promote hematopoiesis.
    • Bone marrow cells can exert inhibitory effects on granulopoiesis within a regulatory microenvironment.