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Isolation and Differentiation of Human Macrophages

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Hypertension

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1527))

Abstract

Macrophage subtypes display protective or pathogenic activities in vascular lesions. They recognize and engulf modified lipids, accumulate in the arterial intima, contributing to the atherosclerosis development. In the heart, vascular tissues and perivascular adipose tissues, there is increasing evidence that macrophages play a role in endothelial dysfunction, vascular inflammation and remodeling. In this chapter we describe protocols to isolate human monocytes from peripheral blood mononuclear cell and how to differentiate them into macrophages by using growth factors (granulocyte-macrophage colony-stimulating factor—GM-CSF or colony-stimulating factor—M-CSF) or by different cell culture medium concentration. We also describe how to indentify different macrophage activation phenotypes.

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References

  1. Auffray C, Sieweke MH, Geissmann F (2009) Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annu Rev Immunol 27:669–692

    Article  CAS  PubMed  Google Scholar 

  2. Bouhlel MA, Derudas B, Rigamonti E, Dievart R, Brozek J, Haulon S et al (2007) PPARgamma activation primes human monocytes into alternative M2 macrophages with anti-inflammatory properties. Cell Metab 6(2):137–143

    Article  CAS  PubMed  Google Scholar 

  3. Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8(12):958–969

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Yan ZQ, Hansson GK (2007) Innate immunity, macrophage activation, and atherosclerosis. Immunol Rev 219:187–203

    Article  CAS  PubMed  Google Scholar 

  5. Koppaka S, Kehlenbrink S, Carey M, Li W, Sanchez E, Lee DE et al (2013) Reduced adipose tissue macrophage content is associated with improved insulin sensitivity in thiazolidinedione-treated diabetic humans. Diabetes 62(6):1843–1854

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Savai R, Pullamsetti SS, Kolbe J, Bieniek E, Voswinckel R, Fink L et al (2012) Immune and inflammatory cell involvement in the pathology of idiopathic pulmonary arterial hypertension. Am J Respir Crit Care Med 186(9):897–908

    Article  CAS  PubMed  Google Scholar 

  7. Sierra-Filardi E, Vega MA, Sanchez-Mateos P, Corbi AL, Puig-Kroger A (2010) Heme oxygenase-1 expression in M-CSF-polarized M2 macrophages contributes to LPS-induced IL-10 release. Immunobiology 215(9-10):788–795

    Article  CAS  PubMed  Google Scholar 

  8. Martinez FO, Gordon S, Locati M, Mantovani A (2006) Transcriptional profiling of the human monocyte-to-macrophage differentiation and polarization: new molecules and patterns of gene expression. J Immunol 177(10):7303–7311

    Article  CAS  PubMed  Google Scholar 

  9. Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M (2004) The chemokine system in diverse forms of macrophage activation and polarization. Trends Immunol 25(12):677–686

    Article  CAS  PubMed  Google Scholar 

  10. Mantovani A, Sica A, Locati M (2005) Macrophage polarization comes of age. Immunity 23(4):344–346

    Article  CAS  PubMed  Google Scholar 

  11. Martinez FO, Helming L, Gordon S (2009) Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol 27:451–483

    Article  CAS  PubMed  Google Scholar 

  12. Sica A, Mantovani A (2012) Macrophage plasticity and polarization: in vivo veritas. J Clin Invest 122(3):787–795

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Francisco J. Rios Ph.D. .

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Rios, F.J., Touyz, R.M., Montezano, A.C. (2017). Isolation and Differentiation of Human Macrophages. In: Touyz, R., Schiffrin, E. (eds) Hypertension. Methods in Molecular Biology, vol 1527. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6625-7_24

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  • DOI: https://doi.org/10.1007/978-1-4939-6625-7_24

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6623-3

  • Online ISBN: 978-1-4939-6625-7

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