Immunocytochemistry of intermediate filaments in cultured arterial smooth muscle cells: Differences in desmin and vimentin expression related to cell of origin and/or plating time

Abstract

The objective of this study was to determine whether intermediate filament expression, including desmin and vimentin, in cultured smooth muscle cells (SMCs) is related to cytodifferentiation or proliferation. Using antibodies to desmin and vimentin, we studied by immunoperoxidase technique the distribution of these proteins in subcultured SMCs derived from porcine aorta and coronary artery. In addition, the proliferative potentiality of the cells was estimated by the incorporation of [ 3H]thymidine into DNA. The frequency of desminpositive cells in coronary arterial SMCs of 3 and 6 population doubling levels was significantly higher as compared to findings with the aortic SMCs and depended on the plating time. No difference was evident at the 12 population doubling level. In contrast, vimentin was present in the majority of both aortic and coronary arterial SMCs. With regard to the localization of vimentin, two cell types were observed, one had reaction products to vimentin in both perinuclear and cell-peripheral areas (type-I cell), the other only in the cell-peripheral region (type-II cell). The relative proportion of the type-I and -II cells varied with the period of culture. Most of the SMCs showed the type-I cell on the first day and the number of type-II cells was increased on the sixth day. Quiescent SMCs in serum-free media had the same percentage of desmin-positive cells and frequency distribution of type-I and -II cells as did the proliferating SMCs incubated in media containing 5% serum. These results suggest that intermediate filament expression, including desmin and vimentin in cultured SMCs, is related to cell origin and/or plating time, but not to the proliferating activity, per se.