Hypertension pp 311-320 | Cite as

Isolation and Differentiation of Human Macrophages

  • Francisco J. Rios
  • Rhian M. Touyz
  • Augusto C. Montezano
Part of the Methods in Molecular Biology book series (MIMB, volume 1527)


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.

Key words

Macrophage Monocyte Human Culture GM-CSF M-CSF 


  1. 1.
    Auffray C, Sieweke MH, Geissmann F (2009) Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annu Rev Immunol 27:669–692CrossRefPubMedGoogle Scholar
  2. 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):137–143CrossRefPubMedGoogle Scholar
  3. 3.
    Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8(12):958–969CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Yan ZQ, Hansson GK (2007) Innate immunity, macrophage activation, and atherosclerosis. Immunol Rev 219:187–203CrossRefPubMedGoogle Scholar
  5. 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):1843–1854CrossRefPubMedPubMedCentralGoogle Scholar
  6. 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):897–908CrossRefPubMedGoogle Scholar
  7. 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):788–795CrossRefPubMedGoogle Scholar
  8. 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):7303–7311CrossRefPubMedGoogle Scholar
  9. 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):677–686CrossRefPubMedGoogle Scholar
  10. 10.
    Mantovani A, Sica A, Locati M (2005) Macrophage polarization comes of age. Immunity 23(4):344–346CrossRefPubMedGoogle Scholar
  11. 11.
    Martinez FO, Helming L, Gordon S (2009) Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol 27:451–483CrossRefPubMedGoogle Scholar
  12. 12.
    Sica A, Mantovani A (2012) Macrophage plasticity and polarization: in vivo veritas. J Clin Invest 122(3):787–795CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Francisco J. Rios
    • 1
  • Rhian M. Touyz
    • 2
    • 1
  • Augusto C. Montezano
    • 1
  1. 1.Institute of Cardiovascular and Medical Sciences, British Heart Foundation Glasgow Cardiovascular Research CentreUniversity of GlasgowGlasgowUK
  2. 2.Kidney Research Centre, Department of Medicine, Ottawa Hospital Research InstituteUniversity of OttawaOttawaCanada

Personalised recommendations