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Umbilical cord endothelial cells expressing large T antigen: Comparison with primary cultures and effect of cell age

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Summary

A number of human endothelial cell lines from umbilical cord cells (HUVECs) have been generated by transfection with SV40 large T and small t antigen sequences. Comparison of these lines with primary cultures of HUVECs has been carried out by monitoring the expression of a number of endothelial cell markers with specific regard to cell age. The secreted levels of the protein plasminogen activator inhibitor (PAI) was found to be significantly reduced in SV40-transfected cells when compared to untransfected controls. Tissue plasminogen activator (tPA) and urokinase (uPA) levels were unchanged. As cells entered crisis, there was a rapid and significant increase in the levels of tPA, uPA, and PAI and this was observed for all clones screened. The endothelial cell marker von Willebrand Factor (vWF) was found intracellularly and was also secreted into the medium. The levels were not altered between transfected and untransfected cells. Angiotensin converting enzyme (ACE) activity was maintained in cell lines at levels found in nonimmortalized HUVECs. Both isoforms (α and β) of IL-1 (interleukin-1) increased as cells approached crisis, and the presence of these cytokines may be responsible for the increased levels of tPA, PAI, and uPA. With one exception, the ability of the transfected cells to produce prostacyclin (PGI2) was lost by all clones.

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Author notes

  1. Una Fitzgerald

    Present address: Department of Neurology and Medical Oncology, University of Glasgow, G12 8QQ, Glasgow, Scotland

Authors and Affiliations

  1. Department of Biological Sciences, University of Paisley, PA1 2BE, Scotland

    Una Fitzgerald, Simon Hettle, Caroline Macdonald & John Stephen McLean

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  1. Una Fitzgerald
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  2. Simon Hettle
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  3. Caroline Macdonald
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  4. John Stephen McLean
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Correspondence to John Stephen McLean.

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Fitzgerald, U., Hettle, S., Macdonald, C. et al. Umbilical cord endothelial cells expressing large T antigen: Comparison with primary cultures and effect of cell age. In Vitro Cell.Dev.Biol.-Animal 36, 222–227 (2000). https://doi.org/10.1290/1071-2690(2000)036<0222:UCECEL>2.0.CO;2

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  • DOI: https://doi.org/10.1290/1071-2690(2000)036<0222:UCECEL>2.0.CO;2

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