Studies on vascular smooth muscle cells and dermal fibroblasts in collagen matrices: Effects of heparin☆

Abstract The incorporation of such tissue-cultured mesenchymal cells as bovine vascular smooth muscle cells (SMS) and human dermal fibroblasts (DF) in a collagen matrix results in the reorganization and distortion of that matrix. A 2 ml collagen matrix populated by 55000 bovine SMC and having a surface area of 800 mm 2 will be reduced to 226 mm 2 by 48 h. Under identical conditions, a lattice populated by 55 000 DF will achieve an area of 78 mm 2 at 48 h. DF are thus more efficient at reducing the size of a collagen lattice by the process of lattice contraction. Bovine SMC proliferate in a collagen matrix; human DF do not. DF in a collagen matrix have a more elongate morphology than SMC. Actin cytoplasmic filaments were studied using the specific F-actin staining reagent, Rhodamine-phalloidin. DF in collagen matrix exhibit diffuse cytoplasmic staining while, in monolayer, they display prominent staining stress fibers. SMC in monolayer and in matrices show stained clumps at the periphery of the cell. The addition of 200 U/ml heparin to SMC eliminates those actin aggregates and causes the formation of stress fibers. It also causes stress fibers to form in dermal fibroblasts in a collagen lattice. However, the elongation and spreading —and the formation of stress fibers brought about by heparin—lead to an inhibition of lattice contraction. Heparin effectively inhibits cell-mediated lattice contraction in SMC and DF, and it also causes the formation of cytoplasmic stress fibers.