Modulation of response to adenosine in vascular smooth muscle cells cultured in defined medium.
Cultured pig aortic smooth muscle cells maintain a viable, quiescent state in a chemically defined medium that contains 10(-6) M insulin, 5 micrograms/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.
Xiong
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YM
Department of Biochemistry, University of Massachusetts, Amherst 01003.
Xu
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SZ
Slakey
L L
LL
D000241:Adenosine / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000166:cytology; D001769:Blood; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D007211:Indoles / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D013552:Swine; D014357:Trypsin / Q000378:metabolism
D003470:Culture Media; D007211:Indoles; C007293:DAPI; D000242:Cyclic AMP; D014357:Trypsin; D000241:Adenosine