Impact of Detachment Methods on M2 Macrophage Phenotype and Function

J Immunol Methods. 2015 Nov:426:56-61. doi: 10.1016/j.jim.2015.08.001. Epub 2015 Aug 5.


The methods of cell detachment influence phenotype and function of human macrophages cultured in vitro. However, comparative studies defining the influence of cell detachment techniques on secondary characterization of M1 or M2 polarized macrophages are largely absent from the literature. In this study we evaluated the impact of trypsin, accutase, EDTA, PBS, and cell scraping on: A. cell recovery, B. phenotype and C. function of in vitro polarized macrophages. Our data demonstrate that while exposure to trypsin or accutase yields highly efficient recovery of viable cells, such chemical cleavage results in loss of select M2 cell surface markers with correlative changes in cell function. In contrast, phenotype and function are maintained following detachment by EDTA on ice. Our data suggest that seemingly "trivial" changes in methodologies for macrophage detachment induce both variable and profound changes on cell phenotype and function which can dramatically impact the results of polarization experiments.

Keywords: CD163; CD206; Macrophage; cell detachment; phenotype.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / biosynthesis
  • Antigens, Differentiation, Myelomonocytic / biosynthesis
  • Biomarkers / metabolism
  • Cell Culture Techniques
  • Cell Separation / methods*
  • Cells, Cultured
  • Collagenases / metabolism*
  • Edetic Acid / metabolism*
  • Humans
  • Lectins, C-Type / biosynthesis
  • Leukocytes, Mononuclear / immunology
  • Lipopolysaccharide Receptors / biosynthesis
  • Macrophages / immunology*
  • Mannose Receptor
  • Mannose-Binding Lectins / biosynthesis
  • Peptide Hydrolases / metabolism*
  • Phenotype
  • Receptors, Cell Surface / biosynthesis
  • Trypsin / metabolism*


  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • Biomarkers
  • CD163 antigen
  • Lectins, C-Type
  • Lipopolysaccharide Receptors
  • Mannose Receptor
  • Mannose-Binding Lectins
  • Receptors, Cell Surface
  • accutase
  • Edetic Acid
  • Peptide Hydrolases
  • Trypsin
  • Collagenases