Freshly isolated femur and tibia from WT C57BL/6 mice were flushed with RPMI-1640 medium (Life Technologies). Cells were harvested and passed through a 40-μm strainer. Red cells were removed with ACK lysis buffer. BM cells were cultured in RPMI-1640 medium supplemented with 5% FBS (Life Technologies). Cells were incubated with 10 ng/ml human CSF-1 (BioLegend, 574808) for 7 days to induce macrophage differentiation. Cells were treated with 100 ng/ml mouse IL-6 (BioLegend, 575708) and CSF-1 for different times.
Human csf 1
Manufactured by BioLegend
Human CSF-1 is a recombinant protein that functions as a cytokine. It is a growth factor that stimulates the proliferation and differentiation of monocytes and macrophages.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using human csf 1
1
Isolation and Differentiation of Murine Bone Marrow Macrophages
2
Monocyte Differentiation Assay
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Primary human monocytes, purchased from the Human Immunology Core at the University of Pennsylvania, were isolated from healthy volunteer donors following leukapheresis by negative selection. All specimens were collected under a University Institutional Review Board-approved protocol, and written informed consent was obtained from each donor. Isolated cells were incubated in RPMI-1640 medium supplemented with 5% heat-inactivated FBS. Cells were treated with 10 ng/ml human CSF-1, 100 ng/ml human IL-6 (BioLegend, 570808), and normal human ECs or patient tumor EC-CM.
3
Monocyte Isolation and Cytokine Treatment
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Primary human monocytes were provided by Human Immunology Core at the University of Pennsylvania. Peripheral blood mononuclear cells were collected from healthy human volunteer donors and monocytes were isolated following leukapheresis by negative selection. All specimens were collected under a University of Pennsylvania Institutional Review Board-approved protocol and written informed consent was obtained from each donor. We have complied with all relevant ethical regulations for work with human participants. Cells were incubated in RPMI-1640 medium supplemented with 5% fetal bovine serum (FBS) and treated with 10 ng/ml human CSF-1 (BioLegend, 574806) and 100 ng/ml human IL-6 (BioLegend, 570808).
4
Monocyte-Macrophage Differentiation and Activation
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Peripheral blood mononuclear cell–derived monocytes were obtained from healthy human volunteer donors ages 16 to 64 of all genders, races, and ethnicities at Human Immunology Core at University of Pennsylvania. Informed consent was obtained from all donors under an Institutional Review Board–approved protocol at University of Pennsylvania. Our work with human participants complies with all relevant ethical regulations. Human primary monocytes were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and treated with human CSF-1 (10 ng/ml; BioLegend, 574806) for 5 days to differentiate into macrophages. Cells were treated with recombinant human OPN (R&D Systems, 1433-OP-050/CF), or cocultured (20:1) with ECs isolated from human GBM tumors, human brain microvascular ECs (ScienCell, 1000), or human glioma cells (U251 cells, Sigma-Aldrich, 09063001; U87 cells, Sigma-Aldrich, 89081402) for 2 days in the presence or absence of or OPN-neutralizing antibody (1 μg/ml; R&D Systems, AF1433-SP).
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