HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell Line

J Invest Dermatol. 1992 Dec;99(6):683-90. doi: 10.1111/1523-1747.ep12613748.

Abstract

The study of human microvascular endothelial cells has been limited, because these cells are difficult to isolate in pure culture, are fastidious in their in vitro growth requirements, and have a very limited lifespan. In order to overcome these difficulties, we have transfected human dermal microvascular endothelial cells (HMEC) with a PBR-322-based plasmid containing the coding region for the simian virus 40 A gene product, large T antigen, and succeeded in immortalizing them. These cells, termed CDC/EU.HMEC-1 (HMEC-1), have been passaged 95 times to date and show no signs of senescence, whereas normal microvascular endothelial cells undergo senescence at passages 8-10. HMEC-1 exhibit typical cobblestone morphology when grown in monolayer culture, express and secrete von Willebrand's Factor, take up acteylated low-density lipoprotein, and rapidly form tubes when cultured on matrigel. HMEC-1 grow to densities three to seven times higher than microvascular endothelial cells and require much less stringent growth medium. HMEC-1 will grow in the absence of human serum, whereas microvascular endothelial cells require culture medium supplemented with 30% human serum. These cells express other cell-surface molecules typically associated with endothelial cells, including CD31 and CD36 and epitopes identified by monoclonal antibodies EN4 and PAL-E. They also express the cell adhesion molecules ICAM-1 and CD44 and following stimulation with interferon-gamma express major histocompatibility complex class II antigens. HMEC-1 specifically bind lymphocytes in cell adhesion assays. Thus HMEC-1 is the first immortalized human microvascular endothelial cell line that retains the morphologic, phenotypic, and functional characteristics of normal human microvascular endothelial cells.

MeSH terms

  • Antigens, Differentiation, Myelomonocytic / analysis
  • Antigens, Polyomavirus Transforming / genetics
  • Cell Adhesion
  • Cell Adhesion Molecules / analysis
  • Cell Adhesion Molecules / genetics
  • Cell Line
  • Endothelium, Vascular / chemistry
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / immunology
  • Humans
  • Intercellular Adhesion Molecule-1
  • Male
  • Microcirculation
  • Phenotype
  • Platelet Endothelial Cell Adhesion Molecule-1
  • T-Lymphocytes / chemistry
  • T-Lymphocytes / cytology
  • Transfection

Substances

  • Antigens, Differentiation, Myelomonocytic
  • Antigens, Polyomavirus Transforming
  • Cell Adhesion Molecules
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Intercellular Adhesion Molecule-1