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Molecular Biotechnology

, Volume 26, Issue 1, pp 7–16 | Cite as

Variations in transfection efficiency of VEGF165 and VEGF121-cDNA

Its effects on proliferation and migration of human endothelial cells
  • Srdjan Djurovic
  • Heidi Kristansen
  • Marit Sletten
  • Kathrine Halvorsen
  • Kåre Berg
Research

Abstract

Little is known about the expression pattern of vascular endothelial growth factor (VEGF) among smooth muscle cells of different arterial regions. Therefore, we have conducted studies aimed at increasing expression of VEGF in cultured human smooth muscle cells (SMCs) from different sites: aorta, umbilical artery, and coronary artery. Two plasmids harboring human VEGF121 and VEGF165 isoforms, respectively, were constructed and lipotransfected into vascular SMCs, using the Fu-GENE 6. Extensive optimization of transfection conditions were performed prior to this.

Different basal levels of VEGF were observed between cell types: from 0.51–0.95 pg/mL/µg protein in umbilical artery, through 2.32–2.39 pg/mL/µg protein in coronary artery, to 5.45–7.52 pg/mL/µg protein in aortic SMCs. Significant differences in responses to transfection were also observed: The increase in VEGF production was most pronounced in umbilical artery SMCs (e.g., with 4 µg VEGF121-cDNA/in the wells)—an approximate 600-fold as opposed to an 18-fold increase in aortic SMCs and a 29-fold increase in coronary artery SMCs.

In addition, we observed significant increases in proliferation rate of aortic and coronary endothelial cells (ECs), after incubation with conditioned medium from VEGF-transfected SMCs. Observed changes differed in relation to cell origin and isoform.

Index Entries

VEGF transfection efficiency smooth muscle cells Fu-GENE 6 proliferation migration 

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Copyright information

© Humana Press Inc 2004

Authors and Affiliations

  • Srdjan Djurovic
    • 1
    • 2
  • Heidi Kristansen
    • 1
    • 2
  • Marit Sletten
    • 1
    • 2
  • Kathrine Halvorsen
    • 1
    • 2
  • Kåre Berg
    • 1
    • 2
  1. 1.Department of Medical GeneticsUllevaal University HospitalOsloNorway
  2. 2.Institute of Medical GeneticsUniversity of OsloNorway

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