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Constitutive secretion of MMP9 by early-passage cultured human endothelial cells

Jacky Arkell1, Christopher J Jackson

  • 1Sutton Arthritis Research Laboratories, Royal North Shore Hospital, St. Leonards, NSW 2065, Australia.

Cell Biochemistry and Function
|November 19, 2003
PubMed

Insights

Early-passage human umbilical vein endothelial cells (HUVE) constitutively secrete matrix metalloproteinase-9 (MMP9). MMP9 secretion decreases with cell passaging, impacting endothelial cell angiogenic potential.

Area of Science:

  • Cell Biology
  • Biochemistry

Background:

  • Matrix metalloproteinase-9 (MMP9) is crucial for angiogenesis.
  • MMP9 is typically induced by stimuli like PMA in endothelial cells.
  • Constitutive expression of MMP9 in endothelial cells is not well-established.

Purpose of the Study:

  • To investigate the constitutive expression and secretion of MMP9 in human umbilical vein endothelial cells (HUVE) across different passages.
  • To determine the passage-dependency of MMP9 secretion in cultured HUVE.

Main Methods:

  • Primary human umbilical vein endothelial cells (HUVE) at passage 0 (P0), passage 1 (P1), and passage 2 (P2) were cultured.
  • MMP9 secretion was analyzed using zymography and ELISA.
  • Gene expression was confirmed via RT-PCR and cycloheximide inhibition studies.

Main Results:

  • P0 HUVE cells constitutively secreted detectable levels of MMP9.
  • MMP9 secretion significantly decreased with successive passaging (P1 and P2).
  • No detectable MMP9 was found in P2 cells under basal conditions, indicating passage-dependent expression.

Conclusions:

  • Cultured HUVE cells constitutively express and secrete MMP9, but only in early passages (P0).
  • MMP9 secretion is significantly reduced as HUVE cells are passaged, likely due to population doubling.
  • These findings suggest that the angiogenic potential of endothelial cells may be linked to their early-passage MMP9 expression.

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