Masuda T, Ohmi K, Yamaguchi H, Hasegawa K, Sugiyama T, Matsuda Y, Iino M, Nonomura Y
1st Department of Pharmacology, Faculty of Medicine, University of Tokyo, Kyowa Hakko Kogyo Co., Japan.
Mol Cell Biochem. 1999 Jan;190(1-2):99-104.
Recently we have established an aortic smooth muscle cell line, p53LMAC01 obtained from p53 knockout mice. This cell line showed some differentiated properties which were accelerated by 5-azacytidine treatment [1]. In this study, further characterization of p53LMAC01 cell line was investigated according to cell growth and differentiation, and especially focused into the changes of cell feature, actin filaments' formation, and changes of intracellular calcium concentrations to sympathetic nerve transmitter, norepinephrine. While the cell feature was changed from flattened shape to extended form during 4 days, actin filaments were developing, arranging in parallel to longitudinal direction, and gathering under the surface membrane. In 11 days many cells died and detached from substrate, while actin filaments became poor except for the surface membrane in the remained cells. Appearance of calcium response to noradrenalin needed several days after passage as well as a morphological change of the cells for the extended form and development of actin filaments. The calcium response was maintained on 11 days, which coincided with the result that the cells hold actin filaments under the surface membrane. These results suggest that p53LMAC01 cell line maintains several differentiated characters of adult smooth muscle cell and that their expression needs several days after passage.
最近，我们建立了一种主动脉平滑肌细胞系p53LMAC01，它源自p53基因敲除小鼠。该细胞系表现出一些分化特性，5-氮杂胞苷处理可加速这些特性的表达[1]。在本研究中，我们根据细胞生长和分化情况对p53LMAC01细胞系进行了进一步表征，特别关注细胞特征的变化、肌动蛋白丝的形成以及细胞内钙浓度对交感神经递质去甲肾上腺素的反应。在4天内，细胞特征从扁平状变为伸长状，同时肌动蛋白丝开始形成，沿纵向平行排列，并在细胞膜下聚集。11天时，许多细胞死亡并从底物上脱落，而剩余细胞中除细胞膜下外，肌动蛋白丝变得稀少。细胞传代后，对去甲肾上腺素的钙反应以及细胞形态变为伸长状和肌动蛋白丝形成需要数天时间。钙反应在11天时仍存在，这与细胞在细胞膜下保留肌动蛋白丝的结果一致。这些结果表明，p53LMAC01细胞系保留了成年平滑肌细胞的一些分化特征，且这些特征在传代后需要数天才能表达。
