Acceleration of endothelial-like cell differentiation from CD14+ monocytes in vitro

Exp Hematol. 2005 Dec;33(12):1554-63. doi: 10.1016/j.exphem.2005.08.011.


Objective: In vitro differentiation of endothelial cells has potential applications in vascular tissue engineering and cell-based therapy for many diseases. The objective of this study was to develop a new strategy that utilizes cytokines and lipopolysaccharide (LPS) to accelerate endothelial-like cell differentiation from peripheral blood CD14(+) monocytes.

Methods: Peripheral blood CD14(+) monocytes were purified with immunobeads and cultured with an angiogenic growth factor-rich growth medium (EGM-2) with or without initial treatment of LPS in combination of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 4 days (the day 4 cultures). The cells were then continuously cultured in EGM-2 medium for an additional 4 or 10 days (the day 8 or day 14 cultures). Cell markers were determined by flow cytometry analysis and immunofluorescence staining. Cytokine/chemokine profile was studied by Bio-Plex immunoassay.

Results: In the group of initial treatment of LPS in combination with GM-CSF, IL-4, and EGM-2, the majority of suspended CD14(+) monocytes were attached and changed their morphology to endothelial-like cells, which expressed high levels of endothelial cell markers CD31, von Willebrand factor, and vascular endothelial growth factor receptor-1 as well as two major endothelial tight junction proteins zonula occludens -1 and occludin in the day 8 cultures. Endothelial nitric oxide synthase expression was substantially increased. Endothelial-like cells were also able to uptake acetylated low-density lipoprotein and bind to Ulex europeus lectin. In addition, endothelial-like cells showed a unique cytokine/chemokine profile with substantial increases of macrophage inflammatory protein-1beta, IL-6, granulocyte colony-stimulating factor, and IL-8.

Conclusion: Initial treatment of LPS in combination with GM-CSF, IL-4, and EGM-2 is an effective strategy for acceleration of endothelial-like cell differentiation from peripheral blood CD14(+) monocytes in vitro.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers / analysis
  • Blood Cells
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects*
  • Cell Shape / drug effects
  • Culture Media / pharmacology
  • Cytokines / biosynthesis
  • Cytokines / pharmacology
  • Endothelial Cells / cytology*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Interleukin-4 / pharmacology
  • Lipopolysaccharide Receptors*
  • Lipopolysaccharides / pharmacology
  • Monocytes / cytology*
  • Monocytes / drug effects
  • Tight Junctions


  • Biomarkers
  • Culture Media
  • Cytokines
  • Lipopolysaccharide Receptors
  • Lipopolysaccharides
  • Interleukin-4
  • Granulocyte-Macrophage Colony-Stimulating Factor