Amédée T, Mironneau C, Mironneau J
Br J Pharmacol. 1986 Aug;88(4):873-80. doi: 10.1111/j.1476-5381.1986.tb16261.x.
A modified method for enzymatically isolating myometrial cells from the pregnant rat has been developed and the mechanical properties of single cells in short-term primary culture have been studied in response to various stimuli. The dissociation method produced a high proportion of fully relaxed cells and these cells shortened and subsequently relaxed completely in response to successive applications of acetylcholine, angiotensin II, high K+ solution or depolarizing current. In single cells, the contractions induced by acetylcholine and high K+ solution were concentration-dependent. Maximal contractions were obtained with 135.6 mM K+ and 5 X 10(-4)M acetylcholine. In single myometrial cells, the time course of contractions induced by acetylcholine, high K+ solution or depolarizing current was similar, suggesting that the rate of shortening was determined by limits of the contractile mechanism. Scanning electron microscopy revealed a smooth surface to the relaxed cells which contrasted with the numerous evaginations present on fully contracted cells. These results demonstrate the retention of structural integrity, acetylcholine and angiotensin II receptors, and potential-dependent Ca channels in myometrial single cells in short-term primary culture. Cells produced by this technique may provide a useful model for detailed electrophysiological studies.
已开发出一种从妊娠大鼠子宫肌层酶解分离细胞的改良方法,并研究了短期原代培养的单细胞在各种刺激下的机械特性。解离方法产生了高比例的完全松弛细胞,这些细胞在连续施加乙酰胆碱、血管紧张素II、高钾溶液或去极化电流时会缩短,随后又完全松弛。在单细胞中,乙酰胆碱和高钾溶液诱导的收缩呈浓度依赖性。在135.6 mM钾离子和5×10⁻⁴M乙酰胆碱时可获得最大收缩。在单个子宫肌层细胞中,乙酰胆碱、高钾溶液或去极化电流诱导的收缩时间进程相似,这表明缩短速率由收缩机制的极限决定。扫描电子显微镜显示,松弛细胞表面光滑,这与完全收缩细胞上存在的大量突起形成对比。这些结果表明,短期原代培养的子宫肌层单细胞保留了结构完整性、乙酰胆碱和血管紧张素II受体以及电压依赖性钙通道。通过该技术产生的细胞可能为详细的电生理研究提供有用的模型。