Open AccessPorcine vascular smooth muscle cells immortalized with SV40 ori‐defective DNA: Characteristics of cell growth and collagen synthesis
Author(s) -
Fujimitsu Kazuhiro,
Sakata Noriyuki,
Jimi Shiro,
Takebayashi Shigeo,
Sasaguri Yasuyuki,
Morimatsu Minoru
Publication year - 1993
Publication title -
acta patholigica japonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.73
H-Index - 74
ISSN - 0001-6632
DOI - 10.1111/j.1440-1827.1993.tb01161.x
Subject(s) - hela , cell culture , transfection , microbiology and biotechnology , biology , cell , actin , cell growth , vascular smooth muscle , smooth muscle , biochemistry , genetics , endocrinology
A cell line derived from medial smooth muscle cells (SMC) was established from the porcine coronary artery by transfection with ori‐defective simian virus 40 plasmid DNA (SV40 DNA). The characteristics of transfected cells (SV40‐SMC) such as cell growth, collagen and non‐collagen syntheses were investigated. SV40‐SMC expressed SV40 large T antigen, c‐myc and c‐myb encoded proteins in the nuclei. SV40‐SMC demonstrated a ‘hills and valleys’ ‐like arrangement in overconfluence and actin filaments upon immunofluorescent staining. Under electron microscopic observation, SV4O‐SMC had larger amounts of synthetic organelles and smaller amounts of filament bundles than those of SMC. SV40‐SMC demonstrated three times higher growth activity and 4.4 times greater cellular density than SMC. Smooth muscle cells did not grow in media containing 5% plasma derived serum (PDS) instead of normal serum, whereas SV40‐SMC proliferated in this medium. SV40‐SMC did not grow in soft agar gel, while HeLa S3 cells, a cell line of human cervical carcinoma, formed colonies in this gel. By immunofluorescent (IF) staining, collagen phenotypes I, Ill, IV and V were detected in both SV4O‐SMC and SMC. However protein synthesis including collagen and non‐collagen was higher in SV40‐SMC than in the control sample. It was concluded that SV40‐SMC were a continuous cell line for vascular SMC regarding morphological characteristics, and demonstrated a higher growth activity, with increased collagen and noncollagen syntheses. This cell line is useful for the investigation of atherogenesis in relation to a proliferation of SMC and an accumulation of extracellular matrices in vascular Intima.