Elsevier

Microvascular Research

Volume 64, Issue 3, November 2002, Pages 384-397
Microvascular Research

Regular Article
In Vitro Expression of the Endothelial Phenotype: Comparative Study of Primary Isolated Cells and Cell Lines, Including the Novel Cell Line HPMEC-ST1.6R

https://doi.org/10.1006/mvre.2002.2434Get rights and content

Abstract

Endothelial cell lines are commonly used in in vitro studies to avoid problems associated with the use of primary endothelial cells such as the presence of contaminating cells, the difficulty in obtaining larger numbers of cells, as well as the progressive loss of cell viability and expression of endothelial markers in the course of in vitro propagation. We have analyzed the characteristics defining distinctive endothelial phenotypes in the cell lines EA.hy926, ECV304, EVLC2, HAEND, HMEC-1, ISO-HAS-1 and a cell line recently generated in our laboratory, HPMEC-ST1.6R, and have compared these phenotypes with those found in primary human endothelial cells isolated from umbilical vein (HUVEC), lung (HPMEC), and skin (HDMEC). The analysis revealed significant differences in phenotype expression between primary cells and the cell lines. Constitutive expression of von Willebrand factor, CD31, and CD34 and induced expression of cell adhesion molecules, ICAM-1, VCAM-1, and E-selectin and cytokines, IL-6, IL-8, MCP-1, and GM-CSF on stimulation with proinflammatory stimuli, as well as the uptake of DiI-Ac-LDL and the formation of cord-like structures on Matrigel, were typically observed in the primary cells. However, most cell lines exhibited only a few of these endothelial characteristics. Only HPMEC-ST1.6R exhibited the major constitutive and inducible endothelial cell characteristics and showed an angiogenic response on Matrigel similar to that of primary HPMEC. Thus, HPMEC-ST1.6R will be a valuable in vitro model system in which to study pathomechanisms and angiogenesis of the mature microvascular endothelium in vitro.

References (51)

  • M. Masuzawa et al.

    Establishment of a new murine-phenotypic angiosarcoma cell line (ISOS-1)

    J. Dermatol. Sci.

    (1998)
  • L.T. May et al.

    Interleukin-6 gene expression in human endothelial cells: RNA start sites, multiple IL-6 proteins and inhibition of proliferation

    Biochem. Biophys. Res. Commun.

    (1989)
  • L. Osborn et al.

    Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes

    Cell

    (1989)
  • L. Richard et al.

    A simple immunomagnetic protocol for the selective isolation and long-term culture of human dermal microvascular endothelial cells

    Exp. Cell Res.

    (1998)
  • K.A. Thomas

    Vascular endothelial growth factor, a potent and selective angiogenic agent

    J. Biol. Chem.

    (1996)
  • H. Vaziri et al.

    Reconstitution of telomerase activity in normal human cells leads to elongation of telomeres and extended replicative life span

    Curr. Biol.

    (1998)
  • E. Venetsanakos et al.

    Induction of tubulogenesis in telomerase-immortalized human microvascular endothelial cells by glioblastoma cells

    Exp. Cell Res.

    (2002)
  • S.M. Albelda et al.

    EndoCAM: A novel endothelial cell–cell adhesion molecule

    J. Cell Biol.

    (1990)
  • Y. Amo et al.

    Expression of vascular endothelial growth factor in a human hemangiosarcoma cell line (ISO-HAS)

    Arch. Dermatol. Res.

    (2001)
  • J. Bauer et al.

    In vitro model of angiogenesis using a human endothelium-derived permanent cell line: Contributions of induced gene expression, G-proteins, and integrins

    J. Cell Physiol.

    (1992)
  • M.P. Bevilacqua et al.

    Identification of an inducible endothelial–leukocyte adhesion molecule

    Proc. Natl. Acad. Sci. USA

    (1987)
  • A.G. Bodnar et al.

    Extension of life-span by introduction of telomerase gene into normal human cells

    Science

    (1998)
  • C.M. Counter et al.

    Dissociation among in vitro telomerase activity, telomere maintenance, and cellular immortalization

    Proc. Natl. Acad. Sci. USA

    (1998)
  • W.G. Dirks et al.

    ECV304 (endothelial) is really T24 (bladder carcinoma): Cell line cross-contamination at source (letter)

    In Vitro Cell Dev. Biol. Anim.

    (1999)
  • M.L. Dustin et al.

    Lymphocyte function-associated antigen (LFA-1) interacting with intercellular adhesion molecule-1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to culture endothelial cells

    J. Cell Biol.

    (1988)
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