Increased Mitotic Activity in Primary Cultures of Aortic Medial Smooth Muscle Cells after Exposure to Hyperlipemic Serum
The vascular smooth muscle cell has been demonstrated to play an active part in the pathogenesis of mammalian atherosclerosis. For several years now, we have been using primary cultures of aortic medial cells in order to study their response to an isolated atherogenic stimulus, namely, diet induced hypercholesteremia. Morphological and immunohistochemical evidence suggest that these cultures consist almost exclusively of smooth muscle cells. Our method of culturing is to place small explants of aortic media in tissue culture flasks with an appropriate medium, let them attach to the bottom of the flask and after a 5–10 day period, the first cells can be observed growing from this explant. Within 5–6 weeks, this outgrowth reaches a diameter of about 15–18 mm. These cell colonies can be maintained in culture for another 2 months or longer. However, they increase very little during that period, not more than 1–2 mm, and only few mitotic figures can be observed. This plateau is quite characteristic for these cells and it is at this stage that we use the cells for our experiments. The cells are elongated, mostly arranged in parallel fashion, often forming characteristic patterns of multilayered growth. Electron microscopical examination and special stains indicate that there is no loss of contact-inhibition in these areas, but rather that the layers are separated by elastin, collagen, or mucopolysaccharides.
KeywordsLipid Uptake Postprandial Serum Aortic Medium High Lipid Diet Multilayered Growth
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