Driska S P, Porter R
Am J Physiol. 1986 Sep;251(3 Pt 1):C474-81. doi: 10.1152/ajpcell.1986.251.3.C474.
A new method is described for the preparation of viable, elongated smooth muscle cells from the swine carotid artery. Cells were prepared by papain digestion of pressurized arteries in calcium-free solution. After digestion, the arteries were everted, and fine strips were teased from the intimal surface of the media in calcium-free solution, releasing single cells. Viability was assessed by exclusion of trypan blue and by appearance under phase-contrast microscopy. By these criteria, approximately 20% of the isolated cells were viable. The most distinguishing and unexpected characteristic of these cells was their length. Mean length of the relaxed viable cells was 240.4 +/- 47.4 microns (SD, n = 76), which is much longer than previously reported for arterial smooth muscle cells. Calcium (1.6 mM) caused most of the viable cells to contract slightly, and the mean cell length in calcium was 194.4 +/- 57.7 microns. Cells in 1.6 mM calcium contracted substantially in response to 10 microM histamine or the calcium ionophore A23187 (10 microM), demonstrating that histamine receptors and the contractile apparatus were still functional.
本文描述了一种从猪颈动脉制备存活的、细长平滑肌细胞的新方法。通过在无钙溶液中用木瓜蛋白酶消化加压动脉来制备细胞。消化后，将动脉翻转，在无钙溶液中从内膜表面的中膜刮出细条，从而释放出单个细胞。通过台盼蓝排斥法和相差显微镜下的外观来评估细胞活力。根据这些标准，大约20%的分离细胞是存活的。这些细胞最显著且出人意料的特征是它们的长度。松弛的存活细胞的平均长度为240.4±47.4微米（标准差，n = 76），这比先前报道的动脉平滑肌细胞长得多。钙（1.6 mM）使大多数存活细胞轻微收缩，在钙存在下的平均细胞长度为194.4±57.7微米。处于1.6 mM钙中的细胞对10 microM组胺或钙离子载体A23187（10 microM）有显著收缩反应，表明组胺受体和收缩装置仍然具有功能。
