Summary
Cultured pig aortic smooth muscle cells maintain a viable, quiescent state in a chemically defined medium that contains 10−6 M insulin, 5µg/ml transferrin, and 0.2 mM ascorbate. DNA synthesis and DNA content were determined by measuring tritiated thymidine incorporation and DNA-binding to the fluorescent probe 4′,6-diamidino-2-phenylindole, respectively. The majority of the population of cells in defined medium cultures were diploid. Tritiated thymidine uptake in cells in defined medium was one-tenth that observed in cells in fetal bovine serum-containing medium. The study of cellular cyclic AMP level in response to extracellular adenosine stimulation in dividing cells and quiescent cells showed that cells in defined medium had a lower extent of response to adenosine compared to cells cultured in serum-containing medium. Both the cell growth index and the response to adenosine of cells cultured in defined medium were reversible after replacing the medium with 10% fetal bovine serum-containing medium, which suggests that the cells in defined medium were healthy and were capable of modulating cellular metabolism depending on culture conditions.
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References
Anand-Srivastava, M. B.; Franks, D. J.; Cantin, M., et al. Presence of “Ra” and “P” site receptors for adenosine coupled to adenylate cyclases in cultured vascular smooth muscle cells. Biochem. Biophys. Res. Commun. 108:213–219; 1982.
Brooker, G.; Harper, J. F.; Terasaki, W. L., et al. Radioimmunoassay of cyclic AMP and cyclic GMP. Adv. Cyclic Nucleotide Res. 10:1–33; 1979.
Burnstock, G.; Buckley, N. J. The classification of receptors for adenosine and adenine nucleotide. Methods Pharmacol. 6:193–211; 1985.
Campbell, J. H.; Campbell, G. R., eds. Vascular smooth muscle in culture. Boca Raton, FL: CRC Press; 1987.
Dickinson, E. S.; Goldman, S. J.; Gordon, E. L., et al. A micromethod for the quantitation by radioimmunoassay of cyclic AMP in samples containing immuno-cross reactive compounds and other interfering substances. J. Cyclic Nucleotide Protein Phosphorylation Res. 11:383–394; 1987.
Goldberg, I. D.; Rosen, E. M. Isolation and culture of a tetraploid subpopulation of smooth muscle cells from the normal rat aorta. Science 226:559–561; 1984.
Goldman, S. J.; Dickinson, E. S.; Slakey, L. L. Effect of adenosine on synthesis and release of cyclic AMP by cultured vascular cells from swine. J. Cyclic Nucleotide Protein Phosphorylation Res. 9:69–78; 1983.
Gordon, D.; Mohai, L. G.; Schwartz, S. M. Induction of polyploidy in cultures of neonatal rat aortic smooth muscle cells. Circ. Res. 59:633–644; 1986.
Gordon, D.; Schwartz, S. M. Replication of arterial smooth muscle cells in hypertension and atherosclerosis. Am. J. Cardiol. 59:44A-48A; 1987.
Holte, H.; Torjesen, P.; Blomhoff, H. K., et al. Cyclic AMP has the ability to influence multiple events during B cell stimulation. Eur. J. Immunol. 18:1359–1366; 1988.
Hu, J.; Olson, E. N. Regulation of differentiation of the BC3H1 muscle cell line through cAMP-dependent and -independent pathways. J. Biol. Chem. 263:19670–19677; 1988.
Jonzon, B.; Nilsson, J.; Fredholm, B. B. Adenosine receptor-mediated changes in cyclic AMP production and DNA synthesis in cultured arterial smooth muscle cells. J. Cell. Physiol. 124:451–456; 1985.
Libby, P.; O’Brien, K. V. Culture of quiescent arterial smooth muscle cells in a defined serum-free medium. J. Cell. Physiol. 115:217–223; 1983.
Linderman, J. J.; Gross, D. J.; Harris, L. J., et al. Charge-coupled device imaging of rapid calcium transients in smooth muscle cells. Cell Calcium. 11:131–144; 1990.
Mioh, H.; Chen, J. Transforming growth factor-β inhibits cellular adenylate cyclase activity in cultured human arterial endothelial cells. In Vitro Cell. Dev. Biol. 25:101–104; 1989.
Rozengurt, E.; Pardee, A. B. Opposite effects of dibutyryl adenosine 3′:5′ cyclic monophosphate and serum on growth of Chinese hamster cells. J. Cell. Physiol. 80:273–280; 1972.
Schwartz, S. M.; Reidy, M. A. Common mechanisms of proliferation of smooth muscle in atherosclerosis and hypertension. Hum. Pathol. 18:240–247; 1987.
Schwartz, S. M.; Ross, R. Cellular proliferation in atherosclerosis and hypertension. Prog. Cardiovasc. Dis. 26:355–372; 1984.
Staples, R. C.; Gross, D.; Tiburzy, R., et al. Changes in DNA content of nuclei in rust uredospore germlings during the start of differentiation. Exp. Mycol. 8:245–255; 1984.
Owens, G. K.; Loeb, A.; Gordon, D., et al. Expression of smooth muscle-specificα-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation. J. Cell. Biol. 102:343–352; 1986.
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This work was supported in part by National Institutes of Health grants HL31854, HL38130, and RR07048.
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Xiong, Y., Xu, S. & Slakey, L.L. Modulation of response to adenosine in vascular smooth muscle cells cultured in defined medium. In Vitro Cell Dev Biol - Animal 27, 355–362 (1991). https://doi.org/10.1007/BF02630954
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DOI: https://doi.org/10.1007/BF02630954