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  • Title: Rat vascular smooth muscle cells immortalized with SV40 large T antigen possess defined smooth muscle cell characteristics including growth inhibition by heparin.
    Author: Reilly CF.
    Journal: J Cell Physiol; 1990 Feb; 142(2):342-51. PubMed ID: 2154504.
    Abstract:
    Rat aortic smooth muscle cells (SMC) have been established by retroviral delivery of the complementary DNA (cDNA) for the simian virus 40 large T antigen (SV40LT) and examined for SMC phenotypic markers and growth characteristics, including responsiveness to the antiproliferative effects of heparin. The transfected cells (SV40LT-SMC) maintain defined SMC characteristics for more than 215 population doublings (PD) as judged by muscle-specific actin expression and growth inhibition by heparin. SV40LT-SMC greater than 129 PD become transformed while SV40LT-SMC less than 77 PD resemble nontransfected SMC morphologically and are nontumorigenic. SV40LT-SMC apparently release a growth factor which acts in an autocrine fashion, since (1) suramin inhibits SV40LT-SMC proliferation, (2) SV40LT-SMC-conditioned medium (CM) contains mitogenic activity, and (3) SV40LT-SMC CM suppresses the binding of platelet-derived growth factor to SMC. Heparin (10-100 micrograms/ml) is a potent inhibitor of both early (less than 80 PD) and late-passage (greater than 80 PD) SV40LT-SMC proliferation. The antiproliferative effects of heparin are similar to those previously observed for SMC by several criteria; the dose-response inhibition curves are indistinguishable from those obtained with nontransfected cells, other glycosaminoglycans have little effect on SV40LT-SMC growth, the antiproliferative effects of heparin are reversed in the presence of epidermal growth factor, and heparin displays high-affinity saturable binding to SV40LT-SMC. In conclusion, SV40LT-SMC are a continuous line of SMC-like cells that are sensitive to the growth inhibitor, heparin. SV40LT-SMC should facilitate studies of heparin inhibition and may be applicable for the study of other SMC characteristics as well.
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