Human m csf

Manufactured by BioLegend

Sourced in Germany

Human M-CSF is a recombinant cytokine that functions as a growth factor for macrophages. It promotes the survival, proliferation, and differentiation of macrophages.

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Lab products found in correlation

Cd14 microbead, by Miltenyi Biotec (2 mentions) Human il 4, by Thermo Fisher Scientific (2 mentions) Mk 8931, by Selleck Chemicals (2 mentions) Cld 8909 liposomes, by Encapsula Nanosciences (2 mentions) Luck 10g, by Santa Cruz Biotechnology (2 mentions) Human il 3, by BioLegend (2 mentions) Tgf β 200 21, by Thermo Fisher Scientific (2 mentions) Human bfgf, by R&D Systems (2 mentions) Protease inhibitor, by Roche (2 mentions) Phosphatase inhibitor, by Roche (2 mentions) Clodronate, by Encapsula Nanosciences (2 mentions) M csf, by BioLegend (2 mentions) G csf, by BioLegend (1 mentions) Anti cd14 antibody, by BD (1 mentions) Heat inactivated human ab serum, by Thermo Fisher Scientific (1 mentions) Recombinant human il 4, by R&D Systems (1 mentions) Rpmi medium, by Thermo Fisher Scientific (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Bovine serum albumin (bsa), by BioLegend (1 mentions) Accutase solution, by Merck Group (1 mentions)

Spelling variants of this product

M-CSF (148 citations) Recombinant human M-CSF (16 citations) Recombinant human macrophage colony-stimulating factor (M-CSF) (6 citations)

12 protocols using human m csf

Monocyte Differentiation into Macrophages

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Peripheral blood mononuclear cells were isolated from buffy coats from healthy donors (Blutspende Zürich, Zurich, Switzerland) by density gradient centrifugation using Ficoll-paque PLUS (Cat: 17–1440–02; GE Healthcare). The monocyte population was magnetically enriched by negative selection using the Human Monocyte Isolation Kit II (Cat: 130–091–153; MACS, Miltenyi Biotech). The monocyte purity was >95% as confirmed by FACS analysis with an anti-CD14 antibody (BD Biosciences).
Freshly isolated human monocytes were differentiated into macrophages by incubation with either 50 ng/mL of human M-CSF (Cat: 14–8789) or G-CSF (Cat: 578602) (both from BioLegend) in RPMI medium (Cat: 61870–010; Gibco) supplemented with 1% heat-inactivated human AB serum (Cat: 34005100; Invitrogen), for up to 8 d. 20 ng/mL of recombinant human IL-4 (Cat: 204-IL; R&D Systems) was used as a control to induce M2-type macrophage differentiation and anti-human G-CSF (1 μg/mL, clone BVD13–3A5, Cat: 502101; BioLegend) or isotype control (rat IgG1 Cat: 400401) was used to neutralize G-CSF in MDA-MB-231 conditioned medium.

Hollmén M., Karaman S., Schwager S., Lisibach A., Christiansen A.J., Maksimow M., Varga Z., Jalkanen S, & Detmar M. (2015). G-CSF regulates macrophage phenotype and associates with poor overall survival in human triple-negative breast cancer. Oncoimmunology, 5(3), e1115177.

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Differentiation of Primary Human Myeloid Cells

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Primary human monocytes, macrophages, and dendritic cells were cultured in RPMI1640 GlutaMAX medium supplemented with 10% FCS. All media contain 1% penicillin/streptomycin, and the cells were cultured at 37 °C in a 5% CO₂ atmosphere. Sc1o was dissolved in DMSO and further diluted in media (c_stock = 25 mM, maximal DMSO concentration during experiments 0.3% v/v EDTA was from Sigma Aldrich (Schnelldorf, Germany). Human FcR blocking reagent, bovine serum albumin, human CD14 MicroBeads, human GM-CSF, human M-CSF, IL-4, and all antibodies for surface staining except CD197 from BioLegend (Fell, Germany) were from Miltenyi Biotec (Bergisch Gladbach, Germany). Human IL-6, IFN-γ, IL-1β, and TNF-α were from PeproTech (Hamburg, Germany). PGE₂ and Accutase solution were from Merck (Darmstadt, Germany).

Blum L., Ulshöfer T., Henke M., Krieg R., Berneburg I., Geisslinger G., Becker K., Parnham M.J, & Schiffmann S. (2020). The immunomodulatory potential of the arylmethylaminosteroid sc1o. Journal of Molecular Medicine (Berlin, Germany), 99(2), 261-272.

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Effect of Bexmarilimab on Macrophage TNFα

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To evaluate the effect of bexmarilimab on TNFα secretion by primary human macrophages, monocytes were enriched from buffy-coat PBMCs from 7 healthy donors (Finnish Red Cross) with CD14 Microbeads (Miltenyi Biotech) and induced to differentiate into M2 macrophages by a 7-day incubation with 50 ng/mL of human M-CSF (Biolegend) followed by a 48-hour incubation with 100 nmol/L dexamethasone (Merck) and 20 ng/mL of human IL4 (200–04–50UG; Peprotech). The cells were treated with bexmarilimab (0.1–50 μg/mL) or irrelevant isotype control (IgG4; Abzena) for 24 hours as described in the Supplementary Methods. After stimulation of TNFα production with a Toll-like receptor 4 (TLR4) agonist [lipopolysaccharide (LPS) EB-ultrapure, E. coli O111:B4; InvivoGen], TNFα levels were measured with human TNFα ELISA kit (Invitrogen) according to manufacturer's instructions and calculated as a mean from triplicate wells per each condition for each donor.

Hollmén M., Maksimow M., Rannikko J.H., Karvonen M.K., Vainio M., Jalkanen S., Jalkanen M, & Mandelin J. (2022). Nonclinical Characterization of Bexmarilimab, a Clever-1–Targeting Antibody for Supporting Immune Defense Against Cancers. Molecular Cancer Therapeutics, 21(7), 1207-1218.

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MK-8931 Pharmacological Study Protocol

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MK-8931 was purchased from Selleckchem (S8173) or Medkoo (331024). Clodronate (SKU# CLD-8909) liposomes and the Control (SKU# CLD-8910) were purchased from Encapsula NanoSciences. Other reagents used in this study includes D-Luciferin (GoldBio, LUCK-10G), etoposide (Santa Cruz, sc-3512B), 32% paraformaldehyde (Electron Microscopy Sciences, 15714), protease inhibitors (Roche, 04693159001), phosphatase inhibitors (Roche, 04906837001), recombinant human EGF (GoldBio, 1150-04-100), human M-CSF (Biolegend, 574806), human IL-3 (Biolegend, 578006), and human bFGF (R&D Systems, 4114-TC-01M). Recombinant human SCF (300-07), VEGF (100-20), IL-4 (200-04), IL-6 (200-06), IL-10 (200-10), sIL-6R (200-06RC), and TGF-β (200-21) were purchased from Peprotech. The sources of other reagents are indicated in experiments described below.

Zhai K., Huang Z., Huang Q., Tao W., Fang X., Zhang A., Li X., Stark G.R., Hamilton T.A, & Bao S. (2021). Pharmacological Inhibition of BACE1 Suppresses Glioblastoma Growth by Stimulating Macrophage Phagocytosis of Tumor Cells. Nature cancer, 2(11), 1136-1151.

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Generating Mouse and Human Macrophages

Cited 1 time

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Mouse bone marrow derived macrophages (BMDM) were generated by culturing bone marrow cells with macrophage colony stimulating factor (MCSF) (BioLegend, 576406) as previously described (47 (link)). Human monocyte derived macrophages (HMDM) were generated by culturing human monocytes with human MCSF (BioLegend, 574804). More details can be found in the supplemental materials.

Mirji G., Worth A., Bhat S.A., Sayed M.E., Kannan T., Goldman A.R., Tang H.Y., Liu Q., Auslander N., Dang C.V., Abdel-Mohsen M., Kossenkov A., Stanger B.Z, & Shinde R.S. (2022). The microbiome-derived metabolite TMAO drives immune activation and boosts responses to immune checkpoint blockade in pancreatic cancer. Science immunology, 7(75), eabn0704.

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Monocyte-Derived M2c Macrophage Polarization

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Monocytes were isolated from whole blood of healthy donors by Ficoll separation and magnetic bead depletion using the RosetteSep Human Monocyte Enrichment Cocktail (Stemcell Technologies, catalog no. 15028). A total of 4 × 10⁵ monocytes per well were seeded in Iscove's modified Dulbecco's medium (IMDM) containing 10% FBS into 24-well plates. For monocyte differentiation, 50 ng/mL human MCSF (BioLegend) was added per well 24 hours after monocyte plating. On day 4, half the media was aspirated and 500 µL fresh media containing 50 ng/mL human MCSF was added per well. To polarize the differentiated M2-like cells to M2c macrophages, on day 6 the entire media content was aspirated and 1 mL of media (IMDM + 10% FBS) containing 50 ng/mL MCSF and 10 ng/mL IL10 (BioLegend) were added per well. On day 7, M2c macrophages were preincubated with anti-PSGL-1 antibody or isotype control antibody at a final concentration of 10 µg/mL for 30 minutes at 37°C, followed by activation with 100 ng/mL LPS (Invivogen) for an additional 24 hours. Cell culture supernatants were analyzed for secreted factors using ELISAs and Luminex.

Kauffman K., Manfra D., Nowakowska D., Zafari M., Nguyen P.A., Phennicie R., Vollmann E.H., O'Nuallain B., Basinski S., Komoroski V., Rooney K., Culyba E.K., Wahle J., Ries C., Brehm M., Sazinsky S., Feldman I, & Novobrantseva T.I. (2023). PSGL-1 Blockade Induces Classical Activation of Human Tumor-associated Macrophages. Cancer Research Communications, 3(10), 2182-2194.

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Isolation and Profiling of Human Monocytes

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Human monocytes were isolated from human buffy coats purchased from the Massachusetts General Hospital blood bank using a standard Ficoll gradient and subsequent CD14+ cell positive selection (Stemcell Technologies). Selected monocytes were cultured in ultra low- adherence flasks (Corning) for 0, 3, or 6 days with RPMI media (Invitrogen) supplemented with 10% FBS (Invitrogen) and 50 ng/mL human M-CSF (Biolegend). SeqWell analysis was performed as previously described⁵⁴. Briefly, at the respective timepoint, cells were detached using trypsin, spun down, and counted. Approximately 12,000 cells were loaded on each array for each timepoint and condition to minimize doublet-loading. The arrays were sealed with a semi-permeable membrane prior to cell lysis and hybridization to single-cell beads. Beads were subsequently pooled for reverse transcription and whole transcriptome amplification.

Hie B., Bryson B, & Berger B. (2019). Efficient integration of heterogeneous single-cell transcriptomes using Scanorama. Nature biotechnology, 37(6), 685-691.

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Monocyte-Derived M2 Macrophage Polarization

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Monocytes were enriched from buffy-coat PBMCs from healthy donors with CD14 Microbeads (Miltenyi Biotec). Monocytes were cultured in Iscove's modified Dulbecco's medium (IMDM; 21980–032; Thermo Fisher) supplemented with 10% FBS, penicillin/streptomycin, and 50 ng/mL human M-CSF (574806; BioLegend) and incubated for 7 days with one medium change. After differentiation, the macrophages were M2-polarized with 100 nmol/L dexamethasone (D-2915; Merck) and 20 ng/mL of human IL-4 (200–04–50UG; PeproTech) for 2 days.

Virtakoivu R., Rannikko J.H., Viitala M., Vaura F., Takeda A., Lönnberg T., Koivunen J., Jaakkola P., Pasanen A., Shetty S., de Jonge M.J., Robbrecht D., Ma Y.T., Skyttä T., Minchom A., Jalkanen S., Karvonen M.K., Mandelin J., Bono P, & Hollmén M. (2021). Systemic Blockade of Clever-1 Elicits Lymphocyte Activation Alongside Checkpoint Molecule Downregulation in Patients with Solid Tumors: Results from a Phase I/II Clinical Trial. Clinical Cancer Research, 27(15), 4205-4220.

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Differentiation of Monocyte-Derived Macrophages

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Human monocyte-derived macrophage: Peripheral blood mononuclear cells (PBMCs) were isolated from leukopacks using standard ficoll procedures. Monocytes were separated from PBMCs via plastic adherence. Monocytes were differentiated into macrophage with 10ng/mL human M-CSF (Biolegend) in RPMI (Gibco) media with 10% FBS for 7 days prior to experimental use. Human primary fibroblasts from MVK-deficient (Cat# GM12014) or population controls (Cat# ND38530) were obtained from Coriell Institute for Medical Research and cultured under recommended conditions.

York A.G., Williams K.J., Argus J.P., Zhou Q.D., Brar G., Vergnes L., Gray E.E., Zhen A., Wu N.C., Yamada D.H., Cunningham C.R., Tarling E.J., Wilks M.Q., Casero D., Gray D.H., Yu A.K., Wang E.S., Brooks D.G., Sun R., Kitchen S.G., Wu T.T., Reue K., Stetson D.B, & Bensinger S.J. (2015). Limiting cholesterol biosynthetic flux spontaneously engages type I IFN signaling. Cell, 163(7), 1716-1729.

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MK-8931 Pharmacological Study Protocol

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