Isolation and characterization of vascular endothelial cells from murine heart and lung

Methods Mol Biol. 2012;843:147-54. doi: 10.1007/978-1-61779-523-7_14.

Abstract

The formation of blood vessel networks is a fundamental event in vertebrate embryo development. Angiogenesis and vasculogenesis are the essential processes in vascular formation. Endothelial cells play a key role during angiogenesis and vasculogenesis, and cultured vascular endothelial cells provide an indispensable model for exploring the molecular mechanisms of angiogenesis and vasculogenesis. In this chapter, we described a protocol using PECAM-1-coated Dynabeads for the isolation of vascular endothelial cells from mouse heart and lung. This method will provide up to 10(7) endothelial cells with high purity (>85%). The isolated endothelial cells retain their in vivo characteristics, such as the expression of the cell surface markers PECAM-1 and ICAM-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / metabolism
  • Cell Adhesion Molecules / metabolism
  • Cell Movement
  • Cell Separation / methods*
  • Dissection
  • Endothelial Cells / cytology*
  • Endothelial Cells / metabolism
  • Flow Cytometry
  • Gelatin / chemistry
  • Gene Knockout Techniques
  • Lung / cytology*
  • Magnets / chemistry
  • Mice
  • Mice, Transgenic
  • Microspheres
  • Myocardium / cytology*
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Signal Transduction

Substances

  • Antigens, CD
  • Cell Adhesion Molecules
  • ICAM-2 protein, mouse
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Gelatin