Culture of rat mesenteric arteriolar smooth muscle cells: effects of platelet-derived growth factor, angiotensin, and nitric oxide on growth

Abstract

We cultured smooth muscle cells as explants from rat mesenteric arterioles (40-200 microns in diameter) obtained by injecting a suspension of iron oxide intraarterially and magnetically separating the arterioles after collagenase digestion of adventitial tissue. In third-passaged cells we ascertained smooth muscle purity of > 98% by characteristic morphology, contraction responses, and specific immunofluorescence staining. Treatment of growth-arrested (in 0.4% fetal calf serum) cells with platelet-derived growth factor (0.3-7.5 nM) or angiotensin II (0.001-1000 nM) induced 3H-thymidine incorporation and cell proliferation in a dose-dependent manner (P < 0.01). S-nitroso-N acetylpenicillamine (0.05-0.5 mM), a nitric oxide-generating compound, inhibited 10% fetal calf serum-induced 3H-thymidine incorporation (P < 0.05) and cell proliferation (P < 0.01). The antimitogenic effect of S-nitroso-N-acetylpenicillamine was significantly reduced by hemoglobin and potentiated by superoxide dismutase (P < 0.01). In addition to a new technique for culturing mesenteric arteriolar smooth muscle cells, these findings provide evidence that platelet-derived growth factor, angiotensin II, and nitric oxide may be involved in their growth control.