High-resolution mass spectrometric analysis of the secretome from mouse lung endothelial progenitor cells

Angiogenesis. 2011 May;14(2):163-72. doi: 10.1007/s10456-011-9200-x. Epub 2011 Jan 15.

Abstract

Recently, we isolated and characterized resident endothelial progenitor cells from the lungs of adult mice. These cells have a high proliferation potential, are not transformed and can differentiate into blood- and lymph-vascular endothelial cells under in vitro and in vivo conditions. Here we studied the secretome of these cells by nanoflow liquid chromatographic mass spectrometry (LC-MS). For analysis, 3-day conditioned serum-free media were used. We found 133 proteins belonging to the categories of membrane-bound or secreted proteins. Thereby, several of the membrane-bound proteins also existed as released variants. Thirty-five proteins from this group are well known as endothelial cell- or angiogenesis-related proteins. The MS analysis of the secretome was supplemented and confirmed by fluorescence activated cell sorting analyses, ELISA measurements and immunocytological studies of selected proteins. The secretome data presented in this study provides a platform for the in-depth analysis of endothelial progenitor cells and characterizes potential cellular markers and signaling components in hem- and lymphangiogenesis.

MeSH terms

  • Animals
  • Culture Media, Conditioned / chemistry
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Lung / cytology*
  • Mass Spectrometry / methods*
  • Mice
  • Mice, Inbred BALB C
  • Neovascularization, Physiologic
  • Proteome / chemistry
  • Proteome / metabolism*
  • Reproducibility of Results
  • Solubility
  • Stem Cells / cytology
  • Stem Cells / metabolism*
  • Subcellular Fractions / metabolism
  • Vascular Endothelial Growth Factor Receptor-1 / metabolism
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism

Substances

  • Culture Media, Conditioned
  • Proteome
  • Vascular Endothelial Growth Factor Receptor-1
  • Vascular Endothelial Growth Factor Receptor-2