摘要
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。
he rat pulmonary artery cells are thin-walled, of small quantity, and they usually show differentresponses from those in vivo after being cultured sccondarily, therefore it is difficult to detect the intra-ceIlular Ca(2+) concentration of them by using fluorescence spectrophotometer, ACAS(570) can be used todetect the dynamic changes of intracellular Ca(2+) concentration of single smooth muscle cell which mustbe of non-contractile type and adhere to the cultural plate firmly. The primary culture of isolated ratpulmonary artery smooth muscle cells was performed by using 0. 25%Collagenase,0. 0125%Elastase,and 0. 125%Trypsin, more than 95%of the isolated cells were tested as smooth muscle cells by indi-rect fluorescence antibody staining method, The cells cultured by this method can be used to detect thedynamic changes of intracellular Ca(2+) concentration by cofocal laser microscope。
出处
《中国医科大学学报》
CAS
CSCD
1996年第3期221-224,共4页
Journal of China Medical University
基金
国家"八五"科技攻关项目
关键词
肺动脉
平滑肌细胞
荧光抗体染色
细胞培养
rat pulmonary artery
smooth muscle cell culture
indirect fluorescence antibody stain-ing method
