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Selective culture of primate marrow-derived macrophages in medium devoid of protein additives
J M Patel1, P A Keene, F P Ross
1Department of Haematology, School of Pathology, South African Institute for Medical Research, Johannesburg.
Abstract:
Viable cultures of bone marrow-derived macrophages (BMM phi) from a primate source, the baboon, were maintained for up to 4 weeks in culture in the absence of any exogenous protein in the medium. Baboon peripheral blood monocytes, spleen, lung, and liver M phi s or human BMM phi failed to survive for greater than 4 days. The protein-free BMM phi cultures were morphologically distinctive by virtue of the extremely dendritic appearance of the M phi s. In contrast baboon marrow cultured in the presence of fetal calf serum led to the overgrowth of fibroblastoid cells and in the presence of horse serum produced numbers of giant cells or polykaryocytes in addition to M phi s. The BMM phi were capable of nonimmune phagocytosis of yeast particles, expressed Ia antigen on their surfaces (59%), and were positive cytochemically for nonspecific (alpha-naphthyl acetate) esterase, oil red O, and tartrate resistant acid phosphatase. The addition of sera to established protein-free BMM phi cultures induced a rapid change of shape, viz., retraction of the dendritic processes and rounding up of the M phi s apparent within 10 min. This shape change was not induced by the addition of hemopoietic growth factors granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte CSF (G-CSF), macrophage CSF (M-CSF), or interleukin 3 (IL-3), nor could it be inhibited by the calcium channel blocking agent Nifedipine. Low levels of M-CSF activity, assayed by the murine bone marrow proliferation assay, were detected in the supernatant.
Insights
Baboon bone marrow macrophages (BMM phi) survived for 4 weeks in protein-free cultures, unlike other primate macrophages. Protein-free conditions maintained BMM phi morphology and function, crucial for macrophage research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophage cultures are essential for immunological studies.
- Maintaining macrophage viability and function in vitro is challenging.
- Exogenous proteins in culture media can influence macrophage behavior.
Purpose of the Study:
- To establish long-term viable cultures of baboon bone marrow-derived macrophages (BMM phi) in protein-free conditions.
- To characterize the morphology and function of protein-free BMM phi.
- To investigate the effects of sera and growth factors on BMM phi.
Main Methods:
- Culturing baboon bone marrow cells in protein-free medium.
- Assessing cell viability and morphology over 4 weeks.
- Evaluating macrophage functions: phagocytosis, Ia antigen expression, and cytochemical staining.
- Observing morphological changes upon addition of sera and growth factors.
Main Results:
- Baboon BMM phi survived for up to 4 weeks in protein-free medium.
- Protein-free BMM phi exhibited a distinctive dendritic morphology.
- These macrophages demonstrated nonimmune phagocytosis and expressed Ia antigen.
- Addition of sera induced rapid shape changes, unlike growth factors.
Conclusions:
- Protein-free culture conditions support long-term viability and characteristic morphology of baboon BMM phi.
- This method offers a valuable system for studying primate macrophages without serum-induced artifacts.
- Further research into the factors influencing BMM phi behavior in vitro is warranted.