Publication: Differential proliferation of rat aortic and mesenteric smooth muscle cells in culture
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Date
1992
Authors
Waldbillig, David K. ; Pang, Stephen C.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
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-transfemngselenium
(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. These results suggest that SMC from
different arterial origins possess different rates of
proliferation. Differences in the rate of in vitro
proliferation in these cell lines are due to differences
in growth fraction, the number of celis traversing the
cell cycle. The mechanisms underlying these
differential proliferative potentials remain to be
determined.
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