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大鼠肺动脉壁薄,平滑肌细胞数少,经人工继代培养后细胞往往显示与在体内时不同的反应,故用荧光分光光度计检测该处细胞内钙难于实现。共焦激光显微钝可以测试一个细胞内钙浓度的动态变化,要求所测试细胞必须是非收缩细胞,且较牢固地粘附在培养皿上。本法应用0.2%Collagenase,0.0125%E-lastase,0.125%Trypsin分离大鼠肺动脉平滑肛细胞进行原代培养,经间接荧光抗体染色法证明95%以上为平滑肌细胞。此法培养的细胞可用于共焦激光显微镜测试细胞内Ca(2+)等浓度的动态变化。
Rat pulmonary artery thin wall, a small number of smooth muscle cells, the cells after artificial subculture often show different reactions in vivo, so the use of fluorescence spectrophotometer detection of intracellular calcium is difficult to achieve. Confocal Laser Micro-dull can test a dynamic change in intracellular calcium concentration, the test cells must be non-contracting cells, and more firmly adhered to the Petri dish. This method uses 0.2% Collagenase, 0.0125% E-lastase, 0.125% Trypsin rat pulmonary artery smooth isolated primary culture, indirect fluorescent antibody staining showed that more than 95% of smooth muscle cells. The cells cultured in this method can be used for confocal laser microscopy to test the dynamic changes of intracellular Ca (2+) concentration.