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Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags
Published on: September 9, 2014
Generation of macrophages from peripheral blood monocytes in the rhesus monkey
Ann E Rozner1, Svetlana V Dambaeva, Jessica G Drenzek
1Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
Abstract:
Macrophages are found in tissues throughout the body and are important immune cells, however, these tissue macrophages are difficult to collect and study. Therefore, the ability to differentiate macrophages from peripheral blood precursors is an important research tool. Macrophage differentiation has been well studied in humans, but differentiation in the non-human primate is poorly characterized. Using human models is not always feasible for invasive experimental studies and, therefore, developing reliable protocols for the non-human primate model is important. We describe a method to differentiate macrophages in vitro in the rhesus monkey by culturing adherent peripheral blood mononuclear cells for five days in RPMI-1640 supplemented with 1% human serum, M-CSF, and IL-1beta. The resulting cells had a distinct macrophage phenotype, the ability to secrete cytokines in response to LPS, and antigen uptake and processing capabilities.
Insights
Researchers developed a method to differentiate rhesus monkey macrophages from blood cells. This protocol provides a valuable tool for studying immune responses in non-human primates, overcoming challenges with tissue macrophage collection.
Area of Science:
- Immunology
- Cell Biology
- Primate Research
Background:
- Tissue macrophages are crucial immune cells but difficult to access for research.
- Human macrophage differentiation is well-understood, but non-human primate models are poorly characterized.
- Non-human primate models are vital for studies where human models are not feasible.
Purpose of the Study:
- To establish a reliable in vitro method for differentiating macrophages from rhesus monkey peripheral blood.
- To provide a valuable research tool for non-human primate immunology and disease modeling.
Main Methods:
- Peripheral blood mononuclear cells from rhesus monkeys were cultured adherent.
- Cells were cultured for five days in RPMI-1640 with 1% human serum, M-CSF, and IL-1beta.
- The resulting cells were analyzed for macrophage characteristics.
Main Results:
- The differentiated cells exhibited a distinct macrophage phenotype.
- These cells demonstrated the capacity to secrete cytokines upon lipopolysaccharide (LPS) stimulation.
- The cells showed functional capabilities including antigen uptake and processing.
Conclusions:
- A novel and effective protocol for in vitro macrophage differentiation from rhesus monkey blood precursors was established.
- This method yields functional macrophages suitable for immunological research in non-human primates.
- The developed protocol supports the use of rhesus monkeys as a model for studying macrophage biology and immune responses.
