Human granulocyte macrophage colony stimulating factor

Manufactured by R&D Systems

Sourced in United States

Human granulocyte-macrophage colony-stimulating factor is a recombinant protein that stimulates the production and function of granulocytes and macrophages.

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Lab products found in correlation

Human interleukin 4, by R&D Systems (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Anti cd14 microbeads, by Miltenyi Biotec (1 mentions) Gentamycin, by Merck Group (1 mentions) Human il 2, by R&D Systems (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Celltiter 96 aqueous one solution reagent, by Promega (1 mentions)

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Human macrophage colony-stimulating factor (6 citations)

2 protocols using human granulocyte macrophage colony stimulating factor

Generating Monocyte-Derived Dendritic Cells

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For preparation of monocyte-derived dendritic cells, the protocol used in this study was published elsewhere [26 (link)]. Briefly, the peripheral blood mononuclear cells (PBMCs) were isolated from whole blood using Ficoll-Paque Plus (GE Healthcare Bio-Sciences AB, Umea, Sweden) reagent. Further, the monocytes were extracted from PBMCs with anti-CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) via standard density gradient centrifugation. Isolated human monocytes were cultivated in RPMI 1640 medium supplemented with 10% fetal calf serum, 800 U/mL human granulocyte-macrophage colony-stimulating factor (R&D Systems, Minneapolis, MN, USA), and 500 U/mL human interleukin-4 (R&D) for 6 days to trigger differentiation into immature DCs (iDCs). iDCs were confirmed with CD1a, CD40, CD54, HLA-DR, CD80, CD83, CD86, and DC-LAMP cell markers by flow cytometry and morphological characteristics. To induce the transformation of iDCs into mature DCs (mDCs), 0.1 ug/mL lipopolysaccharide (LPS) was added to the RPMI 1640 medium for 1.5 days. mDCs were also confirmed using the above-mentioned cell markers and morphological characteristics. The DC-SIGN expression levels between iDCs and mDCs were measured by flowcytometry.

Yang Z.S., Huang S.W., Wang W.H., Lin C.Y., Wang C.F., Urbina A.N., Thitithanyanont A., Tseng S.P., Lu P.L., Chen Y.H, & Wang S.F. (2021). Identification of Important N-Linked Glycosylation Sites in the Hemagglutinin Protein and Their Functional Impact on DC-SIGN Mediated Avian Influenza H5N1 Infection. International Journal of Molecular Sciences, 22(2), 743.

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Cytokine-Dependent Cell Culture Assay

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Human TF-1 cells (erythroblasts, ATCC CRL-2003) and mouse HT-2 clone A5E cells (IL-2 dependent T lymphocytes, ATCC CRL-1841) were cultured at 37°C with 5% (v/v) CO2 in growth medium containing RPMI 1640 (Sigma), 10 % (v/v) heat-inactivated fetal bovine serum (Sigma), 1X gentamycin (Sigma), and 2 ng/mL human granulocyte-macrophage colony-stimulating factor (R&D Systems) or human IL-2 (R&D Systems), respectively. These cytokines were replaced in the assay medium by mouse IL-5 for the TF-1 cells and mouse IL-4 for the HT-2 cells (0.25 ng/mL and 7.5 ng/mL, respectively, R&D Systems) and added to a serial titration of the Abs. Cells were washed and resuspended at a final cell density of 1.1 × 10⁶ cells/mL for the TF-1 cells or 0.2 × 10⁶ cells/mL for the HT-2 cells. Cells were then added to the assay medium before addition of the CellTiter 96 AQ_ueous One Solution Reagent (Promega). After 3h of incubation, the absorbance was measured (Fig E1, F and G).

Godar M., Deswarte K., Vergote K., Saunders M., de Haard H., Hammad H., Blanchetot C, & Lambrecht B.N. (2018). A bispecific antibody strategy to target multiple type 2 cytokines in asthma. The Journal of allergy and clinical immunology, 142(4), 1185-1193.e4.

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