Granulocyte macrophage colony stimulating factor gm csf

Manufactured by BioLegend

Sourced in United States

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that stimulates the production and differentiation of granulocytes and macrophages from hematopoietic precursor cells.

Automatically generated - may contain errors

Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Interleukin 4 (il 4), by BioLegend (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Hepes, by Thermo Fisher Scientific (2 mentions) Interleukin 4 il 4, by BioLegend (2 mentions) Gm csf, by BioLegend (2 mentions) Red blood cell lysis buffer, by Merck Group (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) 2 mercaptoethanol, by Thermo Fisher Scientific (1 mentions) Camptothecin cpt, by Merck Group (1 mentions) Lymphoprep, by Axis-Shield (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions) α mem medium, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

GM-CSF (148 citations) Recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) (4 citations)

14 protocols using granulocyte macrophage colony stimulating factor gm csf

Evaluating Immunostimulant Effects on Dendritic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

B16.F10, B16-OVA, and 4T1-Luci cells were cultured according to the manufacturer’s specifications. DC2.4 cells were cultured in RPMI medium supplemented with 10% FBS and penicillin-streptomycin at 37°C with 5% CO₂. BMDCs were generated from bone marrow cells flushed from the femurs of C57BL/6J mice and were cultured in the DC medium: RPMI 1640 (Gibco) supplemented with 10% FBS (Gibco), sodium pyruvate (Gibco), granulocyte-macrophage colony-stimulating factor (GM-CSF) (20 ng ml⁻¹; BioLegend), interleukin-4 (IL-4) (10 ng ml⁻¹; BioLegend), 2 mM l-glutamine (Gibco), 20 μM 2-mercaptoethanol (Gibco), penicillin-streptomycin (Gibco), 1× nonessential amino acids (Gibco), and 10 mM Hepes (Gibco). The medium was half replaced two times a week. On day 7, nonadherent and loosely adherent cells were the immature BMDCs.
DC2.4 cells and BMDCs were collected and plated in a 24-well plate at 1 million cells per well for 24 hours and were then incubated with PS3D1, PS3D1@DMXAA, or the physical mixture of PS3D1 and DMXAA with indicated dosages for indicated times. Then, the cells were collected and the Ifnb and Cxcl10 mRNA expression levels were analyzed by qPCR.

Liang J., Wang H., Ding W., Huang J., Zhou X., Wang H., Dong X., Li G., Chen E., Zhou F., Fan H., Xia J., Shen B., Cai D., Lan P., Jiang H., Ling J., Cheng Z., Liu X, & Sun J. (2020). Nanoparticle-enhanced chemo-immunotherapy to trigger robust antitumor immunity. Science Advances, 6(35), eabc3646.

+ Open protocol

+ Expand

Characterization of Dendritic Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown in RPMI-1640 or DMEM complete growth media containing Heat-inactivated fetal bovine serum (FBS) (Gibco, USA), and Penicillin-streptomycin solution (Pen/Strep) (HyClone, South Logan, USA). Both Phosphate-buffered saline (PBS) and Hanks’ balanced salt solution (HBSS) were obtained from UFC Biotech (KSA). Lymphoprep™ - 1.077 g/mL was purchased from Axis-Shield PoC AS (Norway). Purified Escherichia coli LPS and Dimethyl Sulfoxide (DMSO)-1.10 g/mL (Sigma-Aldrich®, St. Louis, USA) was used. Vitamin D3 was purchased from Nature Made (USA). All CCR7, CD83, CD80, CD14, CD71 recombinant monoclonal antibodies, recombinant human interleukin 4 (IL-4), and granulocyte-macrophage colony-stimulating factor (GM-CSF) were obtained from BioLegend® (San Diego, California). CD3 was purchased from Invitrogen (Carlsbad, California). Isotype control, CD11c, and CD86 recombinant monoclonal antibodies were purchased from R&D systems (Minneapolis, MN, USA). Lithium Heparin tubes were from Xinle sci&tech co., ltd. (China). Magnesium Sulphate anhydrous (anh. MgSO4) (M.W. =120, 37) was purchased from Panreac Quimica SA, Barcelona, Spain. Camptothecin (CPT) (Sigma Aldrich®, St. Louis, USA) was used.

Aldahlawi A.M., Alzahrani A.T, & Elshal M.F. (2020). Evaluation of immunomodulatory effects of Boswellia sacra essential oil on T-cells and dendritic cells. BMC Complementary Medicine and Therapies, 20, 352.

+ Open protocol

+ Expand

Bone Marrow-Derived Dendritic Cell Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

To obtain BMDCs, the bone marrow was flushed from the femur and tibia, and clusters within the bone marrow suspension were dispersed by vigorous pipetting. After red blood cell (RBC) lysis using RBC lysis buffer, the cells were washed twice with fresh cell culture medium. The cells were seeded with 20 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF) (BioLegend, San Diego, CA, USA) into 100 mm Petri dishes at a concentration of 1 × 10⁶ cells/mL. On days 3 and 6, half of the culture medium was replaced with fresh cell culture medium containing 20 ng/mL GM-CSF. In some experiments, the BMDCs were generated with 10 ng/mL of IL-4 (BioLegend, San Diego, CA, USA) and 20 ng/mL GM-CSF rather than GM-CSF alone. For all experiments, the BMDCs were harvested on day 8. To induce BMDC maturation, the BMDCs were replated in 6- or 24-well plates at 1 × 10⁶ cells/mL in fresh cell culture medium, and 100 ng/mL LPS (Sigma, Burlington, MA, USA) was added for the indicated durations.

Lee H.J., Park J.S., Yoo H.J., Lee H.M., Lee B.C, & Kim J.H. (2020). The Selenoprotein MsrB1 Instructs Dendritic Cells to Induce T-Helper 1 Immune Responses. Antioxidants, 9(10), 1021.

+ Open protocol

+ Expand

Differentiation of Myeloid Dendritic Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were euthanized by overdose of the association of dissociative anesthetic (ketamine, 300 mg/Kg) and alpha 2 adrenoreceptor agonist (xylazine, 30 mg/Kg) administered intraperitoneally. Bone marrow precursors were collected from the tibia and femur, and the cell suspension was adjusted to a concentration of 2 × 10⁵ cells/mL and seeded in a 96-well plate. Cells were maintained for seven days (culture medium was changed every other day) at 37 °C and 5% CO₂ in Iscove’s Modified Dulbecco’s Medium (IMDM), containing 10% fetal bovine serum (FBS), 50 μg/mL gentamicin (Gibco, ThermoFisher Scientific Inc., Waltham, MA, USA) and supplemented with 10 ng/mL Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF; Biolegend, San Diego, CA, USA; #505406) [17 (link)]. This protocol differentiates precursor cells in conventional/myeloid DCs. Supplementary Figure S1 shows the gate strategy and percentages of DCs obtained from precursor cells.

de Mato F.C., Barreto N., Cordeiro G., Munhoz J., Bonfanti A.P., da Rocha-e-Silva T.A., Sutti R., Cruz P.B., Sanches L.R., Bombeiro A.L., Chalbatani G.M., Verinaud L, & Rapôso C. (2023). Isolated Peptide from Spider Venom Modulates Dendritic Cells In Vitro: A Possible Application in Oncoimmunotherapy for Glioblastoma. Cells, 12(7), 1023.

+ Open protocol

+ Expand

Murine Bone Marrow Dendritic Cell Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow from 6-week-old male C57BL/6 mice was used. The femur and fibula were collected and washed with RPMI-1640 medium to obtain cell suspensions. Differentiation of isolated bone marrow cells into dendritic cells was performed as described previously (Sakaki et al., 2013 (link)). Granulocyte-macrophage colony stimulating factor (GM-CSF), a differentiation inducer, was purchased from BioLegend (San Diego, CA, United States).

Takenaka Y., Tanaka R., Kitabatake K., Kuramochi K., Aoki S, & Tsukimoto M. (2022). Profiling Differential Effects of 5 Selective Serotonin Reuptake Inhibitors on TLRs-Dependent and -Independent IL-6 Production in Immune Cells Identifies Fluoxetine as Preferred Anti-Inflammatory Drug Candidate. Frontiers in Pharmacology, 13, 874375.

+ Open protocol

+ Expand

Isolation and Culture of Bone Marrow-Derived Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For bone marrow-derived MC (BMMC) culture, WT bone marrow was obtained from WT C57BF/6 mice in ice-cold Hank's Balanced Salt Solution (HBSS, Gibco), and cultured in complete RPMI containing 10% fetal bovine serum (FBS) (Hyclone), 1 mM nonessential amino acids, 25 mM HEPES, 2 mM L-glutamine, 1 mM sodium pyruvate, 1× Antibiotic-Antimycotic (all reagents from Gibco), 10 ng/ml SCF and 5 ng/ml IL-3 (BioLegend) for 8 to 12 weeks. For BMDC culture, bone marrow was obtained from WT C57BL/6 or CD301b-DTR-GFP mice and cultured in RPMI media containing 10% FBS, 1× GlutaMAX® (Gibco), 20 ng/ml granulocyte macrophage colony-stimulating factor (GM-CSF) (BioLegend), or 20 ng/ml GM-CSF and 50 ng/ml IL-4 for 6-7 days. For bone marrow-derived macrophage (BMM) culture, the same procedures were followed replacing the growth factors with 20 ng/ml macrophage colony-stimulating factor (M-CSF). The rat MC RBL-2H3 cells (ATCC) were cultured in minimum essential medium (MEM) medium (Gibco) containing 15% FBS and antibiotics. JAWSII cells (ATCC) were maintained in MEM α medium (Gibco) supplemented with 20% FBS, 1 mM sodium pyruvate, 100 U/ml penicillin, and 100 μg/ml streptomycin. All cells were cultured at 37 °C in a humidified water-jacketed incubator under 5% CO₂ / 95% air atmosphere.

Choi H.W., Suwanpradid J., Kim I.H., Staats H.F., Haniffa M., MacLeod A.S, & Abraham S.N. (2018). Perivascular Dendritic Cells Elicit Anaphylaxis by Relaying Allergens to Mast Cells via Microvesicles. Science (New York, N.Y.), 362(6415), eaao0666.

+ Open protocol

+ Expand

Generation and Activation of Bone Marrow-Derived Dendritic Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

BMDCs were generated and activated as previously described.²¹ ^,⁴⁴ (link) In short, the femur and tibia were removed from either WT or TLR2-/- mice with the bone marrow being removed in a sterile hood. Red blood cells were lysed and 5 × 10⁵ progenitor cells/mL were resuspended and plated in RPMI 1640 supplemented with 10% FBS and 10 ng/mL Granulocyte-Macrophage Colony Stimulating Factor GM-CSF (Biolegend, San Diego, CA). BMDCs were cultured for 6 days at 37°C in 5% CO₂ with replenishing of complete media and cytokines on days 2 and 4. On day 6, BMDCs were removed and replated at 5 × 10⁵ cells/well. After 2 hours, BMDCs were incubated with a concentration of 10:1 BC, LGG, heat-killed LGG, or HBSS for 4 hours. At 4 hours, 200 ug/mL gentamycin was added to kill all extracellular bacteria and cells were washed 3 times with HBSS. Fresh media then was given and BMDCs were incubated for an additional 20 hours. BMDCs then were stained and analyzed by FCM for expression of CD80 and CD86.

Owens J.A., Saeedi B.J., Naudin C.R., Hunter-Chang S., Barbian M.E., Eboka R.U., Askew L., Darby T.M., Robinson B.S, & Jones R.M. (2021). Lactobacillus rhamnosus GG Orchestrates an Antitumor Immune Response. Cellular and Molecular Gastroenterology and Hepatology, 12(4), 1311-1327.

+ Open protocol

+ Expand

Isolation and Culture of Murine Dendritic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The mice were euthanized; tibiae and femurs were harvested, and the bone marrow cells were collected by flushing the bones with RPMI-1640 using a syringe with a 25-gauge needle and filtering the suspension through a 70-μm-cell strainer to prepare a single-cell suspension. The cells were subsequently incubated with Red Blood Cell (RBC) lysis buffer (Sigma-Aldrich Corp., St. Louis, MO) at 37°C for 5 minutes. The cells were cultured in “complete” RPMI-1640 media supplemented by 20ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF; Biolegend, Inc., San Diego, CA), 5% fetal bovine serum (FBS; Atlanta Biologicals, Inc., Atlanta, GA), 100μg/ml streptomycin and 100U/ml penicillin (Thermo Fisher Scientific Inc, Waltham, MA), 2mM L-glutamine (Sigma-Aldrich Corp., St. Louis, MO) and 1M HEPES (Gibco-Thermo Fisher Scientific Inc, Waltham, MA). On days 3 and 5, the RPMI-1640 medium (supplemented with 20ng/ml GM-CSF) was changed. On day 7, 1×10⁷ DCs were harvested. This culture protocol yields more than 90% CD11c^high monocyte derived DCs.[35 (link)]

Singh R.B., Blanco T., Mittal S.K., Taketani Y., Chauhan S.K., Chen Y, & Dana R. (2020). Pigment Epithelium-derived Factor Secreted by Corneal Epithelial Cells Regulates Dendritic Cell Maturation in Dry Eye Disease. The ocular surface, 18(3), 460-469.

+ Open protocol

+ Expand

Murine Bone Marrow Dendritic Cell Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow cells were collected from the femurs and tibiae of 6- to 8-week-old female C57BL/6 mice and cell suspension was prepared. Cells were incubated with Red blood cell (RBC) lysis buffer (Sigma-Aldrich, St. Louis, MO) at 37°C for 10 minutes. Cells were then seeded in six-well plates at 5×10⁵ cells/well concentration, and were cultured in RPMI-1640 medium supplemented with 20 ng/mL of granulocyte-macrophage colony-stimulating factor (GM-CSF; Biolegend, San Diego, CA), 10% fetal bovine serum (FBS; Atlanta biologicals, Atlanta, GA), 100U/ml penicillin and 100μg/ml streptomycin. On day 3, the culture medium was changed and on day 5, about Π10⁷ bone marrow derived dendritic cells (BMDCs) were harvested.

Tan X., Chen Y., Foulsham W., Amouzegar A., Inomata T., Liu Y., Chauhan S.K, & Dana R. (2018). The Immunoregulatory Role of Corneal Epithelium-Derived Thrombospondin-1 in Dry Eye Disease. The ocular surface, 16(4), 470-477.

+ Open protocol

+ Expand

Murine Bone Marrow-Derived Dendritic Cell Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The bone marrow was flushed out of the euthanized mouse’s femurs and tibias with RPMI 1640. Following tissue disruption and red blood cells lysis, the bone marrow cells were cultured in RMPI 1640 medium supplemented with 50 U/mL penicillin, 50 µg/mL streptomycin, 50 µg/mL gentamycin, 10% v/v fetal calf serum (FBS; all from Gibco, Grand Island, NY, USA), 2 mM L-glutamine (Hyglos GmbH, Bernried, Germany) and 20 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF; Biolegend, San Diego, CA, USA). The medium was partially replenished every day. BMDC were obtained after 7 days of differentiation. These cells were then used as antigen presenting cells (APC) by treating them with 5 µg/mL endotoxin-free OVA in the presence or absence of 5 µg/mL CDA/CDG (InvivoGen, San Diego, CA, USA) or 1 µg/mL lipopolysaccharide (LPS, InvivoGen, San Diego, CA, USA). To elucidate the cross-presentation pathway, BMDC were alternatively treated with 100 nM MG-132, 10 µM lactacystin or 10 µM leupeptin (all from Sigma-Aldrich, Darmstadt, Germany).

Hamouda A.E., Schulze K., Ebensen T., Guzmán C.A, & Lirussi D. (2022). IFN-α/β Signaling Is Required for CDG-Mediated CTL Generation and B Lymphocyte Activation. Pharmaceutics, 14(12), 2821.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!