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Granulocyte macrophage colony stimulating factor

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Granulocyte–macrophage colony-stimulating factor is a protein that regulates the production, differentiation, and function of granulocytes and macrophages. It is a key regulator of immune and inflammatory responses.

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Lab products found in correlation

Penicillin streptomycin, by Merck Group (2 mentions) Matrigel, by Corning (2 mentions) Dimethyl sulfoxide (dmso), by Corning (2 mentions) Intesticult organoid growth medium, by STEMCELL (2 mentions) Mouse mdsc isolation kit, by Miltenyi Biotec (2 mentions) Macrophage colony stimulating factor, by STEMCELL (2 mentions) Celecoxib, by LC Laboratories (2 mentions) Ah23848 hemicalcium salt, by LC Laboratories (2 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by Merck Group (1 mentions) Granulocyte macrophage colony stimulating factor, by Thermo Fisher Scientific (1 mentions) Prostaglandin e2, by Merck Group (1 mentions) Ficoll, by Miltenyi Biotec (1 mentions) Tnf α, by R&D Systems (1 mentions)

3 protocols using granulocyte macrophage colony stimulating factor

1

Organoid Co-culture with MDSCs

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MDCs were isolated from spleen, intestine adenoma of APCMin/+ mice and bone marrow of C57BL/6 from P7 using a Mouse MDSC Isolation Kit (Miltenyi Biotec, 130-094-538) according to the manufacturer’s instructions. Crypt isolation was performed according to the protocol from StemCell Technologies, Inc. (DOCMENT #28223 Version 2.0.1). Crypts (500/well) were then embedded in Matrigel (Corning, growth factor reduced, cat #354230) with MDCs (500 cells/well) at the ratio of 1:1, and cultured in IntestiCult organoid growth medium (StemCell Technologies, Inc., cat #06005) supplemented with 100 μg/ml Penicillin-Streptomycin, 5ng/ml granulocyte–macrophage colony-stimulating factor (Sigma-Aldrich, GF004), 2500 UI/ml macrophage colony-stimulating factor (StemCell Technologies, Inc., cat #78057). COX2 inhibitor (Celecoxib, > 99%) and EP4 inhibitor (AH23848 Hemicalcium salt, ≥ 90%) were purchased from LC Laboratories (cat #c-1502) and Sigma-Aldrich (cat #A8227) respectively, and dissolved in DMSO (Corning, cat #25–950-CQC) as 10mM in stock. Organoids co-cultured with MDSCs were treated with COX2 inhibitor (5–20 μM) and EP4 inhibitor (10–30 μM) from day2 to day14. Changes in organoid morphology after drug therapy were visualized with microscope AxioObserver.Z1with Incubator XLmulti S1.
He X., Smith S.E., Chen S., Li H., Wu D., Meneses-Giles P.I., Wang Y., Hembree M., Yi K., Zhao X., Guo F., Unruh J.R., Maddera L.E., Yu Z., Scott A., Perera A., Wang Y., Zhao C., Bae K., Box A., Haug J.S., Tao F., Hu D., Hansen D.M., Qian P., Saha S., Dixon D., Anant S., Zhang D., Lin E.H., Sun W., Wiedemann L.M, & Li L. (2021). Tumor-initiating stem cell shapes its microenvironment into an immunosuppressive barrier and pro-tumorigenic niche. Cell reports, 36(10), 109674.
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2

Monocyte-Derived Dendritic Cell Generation

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MMDCs were generated as described previously (Bruckner et al., 2012 (link)). In brief, mononuclear cells were isolated from whole blood of human male volunteers between 20 and 30 yr of age by using Ficoll (Miltenyi Biotec) gradient centrifugation and enrichment with anti-CD14 coated MACS beads (Miltenyi Biotec) by following the manufacturer’s instructions. The mononuclear cells were differentiated into immature dendritic cells by culturing in 1 ml RPMI (Gibco) containing 10% FBS (R10), 50 ng IL-4 (PeproTech), and 50 ng granulocyte macrophage colony-stimulating factor (PeproTech) for 6 d in a 24-well format (0.5 ml fresh media was added every 2 d of culture). For maturation, 106 cells were cultured in R10 media containing 50 ng/ml IL-4, 50 ng/ml granulocyte macrophage colony-stimulating factor, 10 µg/ml prostaglandin E2 (Sigma-Aldrich), 20 ng/ml TNFα (R&D Systems), 15 ng/ml IL-6 (PeproTech), and 20 ng/ml IL-1β (PeproTech) for 48 h or with 1 µg/ml lipopolysaccharide (from Salmonella abortus; Sigma-Aldrich) for 24 h.
Brown M., Johnson L.A., Leone D.A., Majek P., Vaahtomeri K., Senfter D., Bukosza N., Schachner H., Asfour G., Langer B., Hauschild R., Parapatics K., Hong Y.K., Bennett K.L., Kain R., Detmar M., Sixt M., Jackson D.G, & Kerjaschki D. (2018). Lymphatic exosomes promote dendritic cell migration along guidance cues. The Journal of Cell Biology, 217(6), 2205-2221.
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3

Organoid Co-culture with MDSCs

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MDCs were isolated from spleen, intestine adenoma of APCMin/+ mice and bone marrow of C57BL/6 from P7 using a Mouse MDSC Isolation Kit (Miltenyi Biotec, 130-094-538) according to the manufacturer’s instructions. Crypt isolation was performed according to the protocol from StemCell Technologies, Inc. (DOCMENT #28223 Version 2.0.1). Crypts (500/well) were then embedded in Matrigel (Corning, growth factor reduced, cat #354230) with MDCs (500 cells/well) at the ratio of 1:1, and cultured in IntestiCult organoid growth medium (StemCell Technologies, Inc., cat #06005) supplemented with 100 μg/ml Penicillin-Streptomycin, 5ng/ml granulocyte–macrophage colony-stimulating factor (Sigma-Aldrich, GF004), 2500 UI/ml macrophage colony-stimulating factor (StemCell Technologies, Inc., cat #78057). COX2 inhibitor (Celecoxib, > 99%) and EP4 inhibitor (AH23848 Hemicalcium salt, ≥ 90%) were purchased from LC Laboratories (cat #c-1502) and Sigma-Aldrich (cat #A8227) respectively, and dissolved in DMSO (Corning, cat #25–950-CQC) as 10mM in stock. Organoids co-cultured with MDSCs were treated with COX2 inhibitor (5–20 μM) and EP4 inhibitor (10–30 μM) from day2 to day14. Changes in organoid morphology after drug therapy were visualized with microscope AxioObserver.Z1with Incubator XLmulti S1.
He X., Smith S.E., Chen S., Li H., Wu D., Meneses-Giles P.I., Wang Y., Hembree M., Yi K., Zhao X., Guo F., Unruh J.R., Maddera L.E., Yu Z., Scott A., Perera A., Wang Y., Zhao C., Bae K., Box A., Haug J.S., Tao F., Hu D., Hansen D.M., Qian P., Saha S., Dixon D., Anant S., Zhang D., Lin E.H., Sun W., Wiedemann L.M, & Li L. (2021). Tumor-initiating stem cell shapes its microenvironment into an immunosuppressive barrier and pro-tumorigenic niche. Cell reports, 36(10), 109674.
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