Differential proliferation of rat aortic and mesenteric smooth muscle cells in culture.
Smooth muscle cells (SMC) from various arterial origins have been successfully maintained in culture. The present study evaluates the proliferative activity of aortic and mesenteric SMC in culture. Aortic and mesenteric SMC were obtained from male Wistar rats by explant and enzyme digestion techniques, respectively. Vascular SMC obtained by either method exhibited a characteristic hill-and-valley growth pattern in culture after confluence and were positively labelled with either anti-smooth muscle actin or myosin by an indirect immunofluorescent method. The rate of incorporation of thymidine into DNA and cell number counting were used as indices of proliferation in vitro. Vascular SMC from passages 4-33 were first synchronized with either Dullbecco's Modified Eagle's Medium (DME) or Ham's F-12 medium, supplemented with insulin-transferring-selenium (ITS), for 72 hours. SMC were then stimulated with 10% bovine serum for either 24 or 72 hours with the former processed for scintillation counting, the latter for cell number determination. The incorporation of tritiated thymidine into DNA following a 2 hour incubation was determined by scintillation counting after perchloric acid extraction. In terms of cell numbers, proliferative responses to bovine serum were determined by Coulter counting. Autoradiography was also carried out in some cultures to determine both thymidine and mitotic labelling indices. The rate of thymidine incorporation in aortic cells was 2-3 fold higher than in mesenteric cells. Aortic and mesenteric SMC lines exhibited similar cell cycle intervals in terms of total duration and individuals cycle parameters. However, the total thymidine index was higher in the aortic than mesenteric SMC.(ABSTRACT TRUNCATED AT 250 WORDS)
Waldbilling
D K
DK
Department of Anatomy, Queen's University, Kingston, Ontario, Canada.
Pang
S C
SC
D000818:Animals; D001011:Aorta / Q000166:cytology; D001158:Arteries / Q000166:cytology; D002453:Cell Cycle; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D008297:Male; D008643:Mesentery / Q000166:cytology; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D051381:Rats; D011919:Rats, Inbred Strains
D003470:Culture Media