Gröschel-Stewart U, Chamley J H, McConnell J D, Burnstock G
Histochemistry. 1975 Jun 5;43(3):215-24. doi: 10.1007/BF00499702.
Immunofluorescent staining with anti-smooth or anti-striated muscle myosin was carried out for 30 minutes at room temperature (18-20 degrees C) on cultures of smooth muscle cells and fibroblasts from guinea-pig vas deferens, taenia coli and ureter, rabbit aorta and chicken gizzard and of cardiac muscle cells and fibroblasts from rat ventricle. With anti-smooth muscle myosin, smooth muscle cells showed an intense fluorescent staining in fine fibrils with an "interrupted" appearance running parallel to the longitudinal axis of the cell throughout the cytoplasm, and also in coarser, "non-interrupted" fibrils (termed here "attachment fibrils") concentrated at the surface of the cell adjacent to the glass coverslip. Fibroblasts in the same cultures showed similar, but much weaker, reactions. When anti-striated myosin was added to the smooth muscle cultures, staining of neither cell type was observed. In contrast, cardiac muscle cells in cultures of rat ventricle did not react anti-smooth muscle myosin, but gave bright fluorescent A-band staining with anti-striated myosin. Fibroblasts in the ventricle cultures were unreactive with anti-striated muscle myosin but gave the characteristic weak reaction with anti-smooth muscle myosin. Thus immunofluorescent stainig with anti-smooth muscle myosin is useful for distinguishing between isolated smooth muscle cells and fibroblasts in tissue culture.
使用抗平滑肌或抗横纹肌肌球蛋白进行免疫荧光染色,在室温(18 - 20摄氏度)下对来自豚鼠输精管、结肠带和输尿管、兔主动脉和鸡砂囊的平滑肌细胞和成纤维细胞,以及来自大鼠心室的心肌细胞和成纤维细胞培养物进行30分钟染色。使用抗平滑肌肌球蛋白时,平滑肌细胞在细纤维中呈现强烈的荧光染色,这些细纤维呈“间断”状,平行于细胞纵轴贯穿整个细胞质,并且在较粗的、“不间断”的纤维(此处称为“附着纤维”)中也有染色,这些纤维集中在细胞与玻璃盖玻片相邻的表面。同一培养物中的成纤维细胞显示出相似但弱得多的反应。当将抗横纹肌肌球蛋白添加到平滑肌培养物中时,未观察到两种细胞类型的染色。相反,大鼠心室培养物中的心肌细胞对抗平滑肌肌球蛋白无反应,但用抗横纹肌肌球蛋白染色时呈现明亮的A带荧光染色。心室培养物中的成纤维细胞对抗横纹肌肌球蛋白无反应,但对抗平滑肌肌球蛋白呈现特征性的弱反应。因此,用抗平滑肌肌球蛋白进行免疫荧光染色有助于在组织培养中区分分离的平滑肌细胞和成纤维细胞。