Porcine vascular smooth muscle cells immortalized with SV40 ori-defective DNA: characteristics of cell growth and collagen synthesis.
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, SV40-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, III, IV and V were detected in both SV40-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 non-collagen 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.
Fujimitsu
K
K
Second Department of Pathology, School of Medicine, Fukuoka University, Japan.
Sakata
N
N
Jimi
S
S
Takebayashi
S
S
Sasaguri
Y
Y
Morimatsu
M
M
D000362:Agar; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000378:metabolism; D002455:Cell Division; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D003114:Colony-Forming Units Assay; D004279:DNA, Viral / Q000235:genetics*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D005455:Fluorescent Antibody Technique; D005782:Gels; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism*; D009857:Oncogenes; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D013552:Swine
D000957:Antigens, Viral, Tumor; D004279:DNA, Viral; D016326:Extracellular Matrix Proteins; D005782:Gels; D000362:Agar; D003094:Collagen