Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood

Mol Biol Rep. 2020 Sep;47(9):6929-6940. doi: 10.1007/s11033-020-05751-6. Epub 2020 Sep 2.

Abstract

Macrophages are among the most important cells of the immune system. Among other functions, they take part in almost all defense actions against foreign bodies and bacteria, being particularly important in infections, wound healing, and foreign body reactions. Considering their importance for the health of the human body, as well as their important role in several diseases, the in vitro studies based on these cells, are a crucial research field. Taking all mentioned into account, this study describes a simple isolation method of human macrophages (MFUM-HMP-001 and MFUM-HMP-002 cell lines) from peripheral blood. For this purpose, the morphology, the viability, and the phagocytotic activity of the isolated cells were tested. The Immunostaining of MFUM-HMP-001 and MFUM-HMP-002 cells confirmed the macrophage cell markers CD68, CD80, and CD163/M130. The phagocytotic activity was marked in both MFUM-HMP-001 and MFUM-HMP-002 cells, as was the phagocytosis of the pHrodo green Escherichia coli bioparticles conjugates, which was enhanced with the addition of lipopolysaccharide. The cells were stable and exhibited good growth. According to our results, both cell lines are useful for the development of novel macrophage cell-based in vitro models.

Keywords: Cell characterization; Cell isolation protocol; Human peripheral blood; Macrophages; Phagocytotic activity.

MeSH terms

  • Antigens, CD / blood
  • Antigens, Differentiation, Myelomonocytic / blood
  • B7-1 Antigen / blood
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Survival / drug effects
  • Cells, Cultured
  • Escherichia coli / metabolism
  • Humans
  • Lipopolysaccharides / pharmacology
  • Macrophages / cytology*
  • Macrophages / drug effects
  • Macrophages / metabolism*
  • Phagocytosis* / drug effects
  • Receptors, Cell Surface / blood

Substances

  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • B7-1 Antigen
  • CD163 antigen
  • CD68 antigen, human
  • CD80 protein, human
  • Lipopolysaccharides
  • Receptors, Cell Surface