Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture
Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture
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成人、猴和兔血管平滑肌细胞原代培养和传代培养的比较
DOI：
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发表时间：
1977
影响因子：
3.6
通讯作者：
U. Gröschel‐Stewart
中科院分区：
文献类型：
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作者：
J. H. Chamley;G. Campbell;J. D. McConnell;U. Gröschel‐Stewart
本文介绍了一种原代培养人、猴和兔血管平滑肌细胞的方法，在培养的最初几天内，这些细胞与体内细胞非常相似，在血管紧张素II、去甲肾上腺素和机械刺激下可诱导收缩。它们被抗平滑肌肌动蛋白和肌球蛋白的抗体强烈染色。成纤维细胞和内皮细胞不被这些抗体染色，从而允许监测每批培养物的纯度。这一直被发现优于99%。培养约9天后，平滑肌细胞发生改变或“去分化”，在形态上类似于成纤维细胞。在这个阶段，细胞不能再被诱导收缩，也不能被肌球蛋白抗体染色。这些细胞的强烈增殖很快导致形成汇合单层，在该阶段一些分化特征恢复。通过存在成纤维细胞或内皮细胞的饲养层，或向培养基中添加cAMP，可以抑制修饰或“去分化”过程。在任何时候都具有“去分化”细胞的形态和功能特性。平滑肌细胞在培养的促有丝分裂剂的研究在动脉粥样硬化进行了讨论。
SummaryA method is presented for growing large numbers of pure isolated smooth muscle cells from adult human, monkey, and rabbit blood vessels in primary culture.In the first few days in culture these cells closely resembled those in vivo and could be induced to contract with angiotensin II, noradrenaline and mechanical stimulation. They stained intensely with antibodies against smooth muscle actin and myosin. Fibroblasts and endothelial cells did not stain with these antibodies thereby allowing the purity of each batch of cultures to be monitored. This was consistently found to be better than 99%. The smooth muscle cells modified or “dedifferentiated” after about 9 days in culture to morphologically resemble fibroblasts. At this stage cells could no longer be induced to contract and did not stain with the myosin antibodies. Intense proliferation of these cells soon resulted in a confluent monolayer being formed at which stage some differentiated characteristics returned. The modification or “dedifferentiation” process could be inhibited by the presence of a feeder layer of fibroblasts or endothelial cells, or the addition of cAMP to the culture medium.Smooth muscle cells which had migrated from explants in primary culture, and cells in subculture, had morphological and functional properties of “dedifferentiated” cells at all times.The advantages of differentiated rather than “dedifferentiated” smooth muscle cells in culture for the study of mitogenic agents in atherosclerosis is discussed.