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Recombinant human M-CSF protein (Qk075)

Macrophage colony-stimulating factor (M-CSF) is a cytokine that regulates the survival, proliferation, differentiation, and functional activation of monocytes, such as macrophages and dendritic cells. Various cells produce and release M-CSF, such as endothelial cells, fibroblasts, osteoblasts, smooth muscle, and macrophages.  M-CSF binds to the CSF-1 receptor (CSF-1R) leading to the activation of the PI3K-AKT and AMPK signaling.

M-CSF has been used in vitro for the differentiation of human induced pluripotent stem cell (iPSC)-derived macrophages as well as the maintenance and development of monocytes in hematopoietic stem cell culture. M-CSF has also been used to influence macrophage polarization towards an anti-inflammatory or M2 macrophage phenotype.

A highly bioactive, animal-origin-free (AOF), carrier protein-free 36.8 kDa recombinant protein with exceptional lot-to-lot consistency.

Orders are typically shipped same or next day (except Friday).
Easy world-wide ordering, direct or through our distributors.

1mg will be despatched as 2 x 500µg

Buy online with secure credit card or purchase order or email orders@qkine.com
Bulk and stock reservation available

Summary

  • High purity human M-CSF protein (Uniprot: P09603)

  • >98%, by SDS-PAGE quantitative densitometry

  • Expressed in E. coli

  • 36 kDa dimer

  • Animal-free (AOF)

  • Carrier protein-free

  • Manufactured in our Cambridge, UK laboratories

  • Lyophilized from acetonitrile/TFA

  • Resuspend in 10 mM HCl, prepare single-use aliquots, add carrier protein if desired and store frozen at -20oC or -80oC

Featured applications

  • Differentiation of iPSC-derived macrophages

  • Culture of bone marrow-derived macrophages

  • Maintenance of macrophages

  • Osteoclast culture

  • Polarization of anti-inflammatory or M2 macrophage

  • Differentiation of alveolar macrophages to study fibrotic disorders

Colony-stimulating factor-1, CSF-1, CSF1, macrophage colony stimulating factor, macrophage colony-stimulating factor 1, M-CSF, MCSF, Lanimostim, Flanimostim, MGC31930

1. Shereck EB, van de Ven C, Cairo MS. Neonatal Leukocyte Physiology and Disorders. Avery’s Diseases of the Newborn. 2012 Jan 1;1108–22.

2. Stanley ER, Cifone M, Heard PM, Defendi V. Factors regulating macrophage production and growth: the identity of colony-stimulating factor and macrophage growth factor. Journal of Experimental Medicine. 1976 Mar 1;143(3):631–47.

3. Sehgal A, Irvine KM, Hume DA. Functions of macrophage colony-stimulating factor (CSF1) in development, homeostasis, and tissue repair. Semin Immunol. 2021 Apr 1;54:101509.

4. Hume DA, MacDonald KPA. Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling. Blood. 2012 Feb 23;119(8):1810–20.

5. Mun SH, Park PSU, Park-Min KH. The M-CSF receptor in osteoclasts and beyond. Experimental & Molecular Medicine 2020 52:8. 2020 Aug 17;52(8):1239–54.

6. Breen FN, Hume DA, Weidemann MJ. Interactions among granulocyte-macrophage colony-stimulating factor, macrophage colony-stimulating factor, and IFN-gamma lead to enhanced proliferation of murine macrophage progenitor cells. The Journal of Immunology. 1991 Sep 1;147(5):1542–7.

7. Caracciolo D, Shirsat N, Wong GG, Lange B, Clark S, Rovera G. Recombinant human macrophage colony-stimulating factor (M-CSF) requires subliminal concentrations of granulocyte/macrophage (GM)-CSF for optimal stimulation of human macrophage colony formation in vitro. Journal of Experimental Medicine. 1987 Dec 1;166(6):1851–60.

8. Shadle PJ, Allen JI, Geier MD, Koths K. Detection of endogenous macrophage colony-stimulating factor (M-CSF) in human blood. Exp Hematol. 1989 Feb;17(2):154–9.

9. Chitu V, Stanley ER. Colony-stimulating factor-1 in immunity and inflammation. Curr Opin Immunol. 2006;18(1):39–48.

10. Pandit J, Bohm A, Jancarik J, Halenbeck R, Koths K, Kim SH. Three-dimensional structure of dimeric human recombinant macrophage colony-stimulating factor. Science (1979). 1992;258(5086):1358–62.

11. Pollard JW. Trophic macrophages in development and disease. Nat Rev Immunol [Internet]. 2009 Apr [cited 2023 May 2];9(4):259–70. Available from: https://pubmed.ncbi.nlm.nih.gov/19282852/

12. Hume DA, MacDonald KPA. Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling. Blood [Internet]. 2012 Feb 23 [cited 2023 May 2];119(8):1810–20. Available from: https://ashpublications.org/blood/article/119/8/1810/30399/Therapeutic-applications-of-macrophage-colony

13. Hamilton JA. Colony-stimulating factors in inflammation and autoimmunity. Nature Reviews Immunology 2008 8:7 [Internet]. 2008 Jul [cited 2023 May 2];8(7):533–44. Available from: https://www.nature.com/articles/nri2356

14. Ushach I, Zlotnik A. Biological role of granulocyte macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) on cells of the myeloid lineage. J Leukoc Biol [Internet]. 2016 Sep 1 [cited 2023 May 2];100(3):481. Available from: /pmc/articles/PMC4982611/

15. Chitu V, Gokhan Ş, Nandi S, Mehler MF, Stanley ER. Emerging Roles for CSF-1 Receptor and its Ligands in the Nervous System. Trends Neurosci. 2016;39(6):378–93.

16. Barca C, Foray C, Hermann S, Herrlinger U, Remory I, Laoui D, et al. The Colony Stimulating Factor-1 Receptor (CSF-1R)-Mediated Regulation of Microglia/Macrophages as a Target for Neurological Disorders (Glioma, Stroke). Front Immunol. 2021;12(December):1–12.

17. Stanley ER, Chitu V. CSF-1 receptor signaling in myeloid cells. Cold Spring Harb Perspect Biol [Internet]. 2014 [cited 2023 May 2];6(6). Available from: https://pubmed.ncbi.nlm.nih.gov/24890514/

18. Cannarile MA, Weisser M, Jacob W, Jegg AM, Ries CH, Rüttinger D. Colony-stimulating factor 1 receptor (CSF1R) inhibitors in cancer therapy. J Immunother Cancer. 2017;5(1):1–13.

19. Stanley ER, Chitu V. CSF-1 receptor signaling in myeloid cells. Cold Spring Harb Perspect Biol. 2014;6(6):1–21.

20. Mossadegh-Keller N, Sarrazin S, Kandalla PK, Espinosa L, Richard Stanley E, Nutt SL, et al. M-CSF instructs myeloid lineage fate in single haematopoietic stem cells. Nature [Internet]. 2013 May 5 [cited 2023 May 2];497(7448):239. Available from: /pmc/articles/PMC3679883/

21. Lachmann N, Ackermann M, Frenzel E, Liebhaber S, Brennig S, Happle C, et al. Large-Scale Hematopoietic Differentiation of Human Induced Pluripotent Stem Cells Provides Granulocytes or Macrophages for Cell Replacement Therapies. Stem Cell Reports [Internet]. 2015 Feb 2 [cited 2023 May 2];4(2):282. Available from: /pmc/articles/PMC4325194/

22. Vogel DYS, Glim JE, Stavenuiter AWD, Breur M, Heijnen P, Amor S, et al. Human macrophage polarization in vitro: maturation and activation methods compared. Immunobiology [Internet]. 2014 [cited 2023 May 4];219(9):695–703. Available from: https://pubmed.ncbi.nlm.nih.gov/24916404/

23. Mia S, Warnecke A, Zhang XM, Malmström V, Harris RA. An optimized protocol for human M2 macrophages using M-CSF and IL-4/IL-10/TGF-β yields a dominant immunosuppressive phenotype. Scand J Immunol [Internet]. 2014 [cited 2023 May 4];79(5):305–14. Available from: https://pubmed.ncbi.nlm.nih.gov/24521472/

24. Lescoat A, Ballerie A, Augagneur Y, Morzadec C, Vernhet L, Fardel O, et al. Distinct Properties of Human M-CSF and GM-CSF Monocyte-Derived Macrophages to Simulate Pathological Lung Conditions In Vitro: Application to Systemic and Inflammatory Disorders with Pulmonary Involvement. Int J Mol Sci [Internet]. 2018 Mar 17 [cited 2023 May 4];19(3). Available from: https://pubmed.ncbi.nlm.nih.gov/29562615/

Bioactivity

Bioactivity graph showing the EC50 = 196 pM (7.2 ng/mL) for Qkine recombinant M-CSF protein

M-CSF activity is determined using the proliferation of NFS-60 mouse myeloid leukemia cells. Cells are treated in triplicate with a serial dilution of M-CSF for 48 hours. Cell viability is measured using the CellTiter-Glo (Promega) luminescence assay and normalized. EC50 = 196 pM (7.2 ng/mL). Data from Qk075 lot #204507.

Purity

Human M-CSF protein purity graph (Qk075)

Recombinant M-CSF migrates as a major band at approximately 18 kDa (monomer) in non-reducing (NR) conditions. The dimeric form is the minor band at the higher molecular weight (36 kDa). Upon reduction (R), only the 18 kDa band is visible. No contaminating protein bands are present.

The purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk075 batch #204507.

Further quality assays

  • Mass spectrometry, single species with expected mass
  • Endotoxin: <0.005 EU/μg protein (below level of detection)

  • Recovery from stock vial: >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Protein background

Macrophage colony-stimulating factor (M-CSF) or colony-stimulating factor-1 (CSF-1), is a secreted glycoprotein belonging to the family of hematopoietic growth factors [1]. It is a cytokine which regulates the survival, proliferation, and renewal of monocytes and macrophages [2–4]. It plays a key role in the differentiation of progenitors to the monocyte-macrophage lineage and the functional activation of mature macrophages [4]. It is also involved in the growth and differentiation of other myeloid cells such as dendritic cells and osteoclasts [5]. M-CSF was the first CSF to be isolated and purified [2]. Other growth factors involved in the regulation of monocyte/macrophage include GM-CSF, IL-3, and IFN-γ [6,7].

Human M-CSF has a molecular weight of 36 kDa [8]. It is expressed as three functionally major isoforms: a secreted chondroitin sulfate proteoglycan, a secreted glycoprotein, and a membrane-spanning cell-surface glycoprotein [9]. All three isoforms share the same amino terminal of 149 amino acids required for biological activity [9,10]. The overall topology of M-CSF is dimeric and composed of two covalently linked four-helix bundles forming a flat and elongated structure [10].

Various cells produce and release M-CSF such as endothelial cells, fibroblasts, osteoblasts, smooth muscle, and macrophages [11–14]. M-CSF binds to CSF-1 receptor (CSF-1R), a receptor tyrosine kinase expressed at the surface of bone marrow-derived macrophages, monocytes, microglial cells, as well as osteoclasts and dendritic cells [15–17]. The M-CSF/CSF-1R binding leads to the activation through dimerization of the PI3K-AKT and AMPK signaling cascades involved in macrophage differentiation [16,18].

Low levels of M-CSF stimulate microglia survival while elevated levels of M-CSF stimulate cell proliferation and migration [16,19]. Elevated levels of M-CSF usually result from inflammatory conditions or from immunosuppressive conditions when the tumour microenvironment recruits tumor-associated macrophages [16]. Hence, the M-CSF/CSF-1R axis has gained attention for its potential as a target for neurological disorders such as glioma or stroke [16].

M-CSF has been used in vitro for the differentiation of human induced pluripotent stem cell (hiPSCs)-derived macrophages as well as the maintenance and development of monocytes in hematopoietic stem cell culture [20,21]. In a pluripotent stem cell-based hematopoietic differentiation model, M-CSF along with IL-3 directed myeloid cells to monocyte/macrophage-type cells able to phagocytose and secrete cytokines [21]. In mouse hematopoietic stem cells, M-CSF can instruct myeloid cell-fate change by inducing the myeloid master regulator PU.1 [20]. M-CSF has also been used to influence macrophage polarization. Indeed, previous publications have reported that M-CSF induces an anti-inflammatory or M2 macrophage phenotype [22,23]. Finally, Lescoat et al. have identified human blood monocyte-derived macrophages differentiated with M-CSF as a relevant in vitro model to study alveolar macrophages in fibrotic disorders [24].

Additional resources

Technote: M-CSF

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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