Isolation of vascular smooth muscle cell cultures with altered responsiveness to the antiproliferative effect of heparin

Journal of Cellular Physiology
B L CalebJ J Castellot

Abstract

Smooth muscle cell (SMC) hyperplasia in the arterial wall is an important component of both atherogenesis and post-vascular surgical restenosis. One naturally-occurring group of molecules which can suppress SMC proliferation in animal models and in cell culture systems are the complex carbohydrates of the heparan sulfate class, including heparin. In this communication, we have used retrovirus vectors to introduce several oncogenes into SMC: SV40 Large T antigen (SVLT), polyoma virus Large T antigen (PyLT), v-myc, and adenovirus E1a. We analyzed a total of 11 cultures. A combination of Western blot analysis, immunoprecipitation, and indirect immunofluorescence confirmed the expression of the infected oncogenic protein in each culture we isolated. All four oncogenes permitted the maintenance of a normal SMC phenotype, as assessed by the general morphology of cells in the light microscope and the presence of SMC-specific alpha-actin in an immunofluorescence assay. Doubling times in infected cells ranged from 20 to 33 hr, and final cell densities in infected cultures ranged from 4 x 10(4) to 5 x 10(5) cells per cm2. By comparison, the parent line had a doubling time of 30 hr and reached a final cell density of 1 x 10(5) cells per c...Continue Reading

References

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Citations

Feb 27, 1999·Journal of Cellular Physiology·K Mishra-Gorur, J J Castellot
Sep 10, 1998·American Journal of Respiratory and Critical Care Medicine·K OkadaM D Botney
Jun 27, 1998·Connective Tissue Research·J D San AntonioL A Pukac
Aug 5, 2009·Journal of Cellular Physiology·Hicham MrabatCharles A Hales
Nov 25, 1997·Journal of Cellular Physiology·L M Delmolino, J J Castellot
Jul 1, 1997·Canadian Journal of Physiology and Pharmacology·A J HalaykoN L Stephens
Jul 30, 2002·The Journal of Biological Chemistry·Ketu Mishra-GorurJohn J Castellot

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