Growth and phenotypic characterization of porcine coronary artery smooth muscle cells |
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Authors: | M C Lavigne P W Ramwell R Clarke |
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Institution: | (1) Department of Physiology and Biophysics, Georgetown University Medical Center, 20007 Washington, DC;(2) Present address: Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, 20892 Bethesda, Maryland;(3) Room W405A, The Research Building, Vincent T. Lombardi Cancer Center, Georgetown University Medical Center, 3970 Reservoir Road N.W., 20007 Washington, DC |
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Abstract: | Summary Vascular smooth muscle cell (VSMC) proliferation significantly contributes to atherosclerotic plaque formation and limits
the success rate of percutaneous transluminal coronary angioplasty. We derived a population of porcine coronary artery SMCs
to characterize VSMC proliferation and phenotype in preparation to study the molecular actions of VSMC mitogens and antiproliferative
agents. Growth assays were designed to minimize the estrogen content in the culture medium, since this steroid hormone significantly
influences VSMC growth and the expression of VSMC mitogens and their receptors. Culture conditions were identified such that
this criterion was achieved while maintaining a significant VSMC growth rate. Cells cultured in serum-free medium, regardless
of growth factor supplements, did not remain adherent to a plastic culture substrate, nor did they proliferate. Dextran-coated
charcoal (DCC)-treated sera, including fetal bovine, calf, and porcine, supported VSMC adhesion, but not growth. Whole fetal
bovine serum (FBS) produced the best proliferative response. A type-I collagen-coated culture surface significantly enhanced
VSMC growth, but only in culture medium containing non-DCC-treated FBS. Flow cytometry analyses confirmed the mitogenic effects
of this substrate. The VSMCs exhibited a morphological change on type-I collagen, but this was not accompanied by a change
in VSMC phenotype. Our data indicate that culture of these porcine coronary artery SMCs in 2.5% FBS plus 10 ng platelet-derived
growth factor-BB per ml in phenol red-free medium on type-I collagen may be the optimal conditions for studying the molecular
aspects of VSMC mitogens and antiproliferative agents. |
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Keywords: | serum estrogen mitogens type-I collagen |
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