期刊论文

Methods of Culture of Rat Pulmonary Artery Smooth Muscle Cells and Identification

中文

中南医学科学杂志

2095-1116

2015

1

78-81

10.15972/j.cnki.43-1509/r.2015.01.020

13C837:湖南省教育厅科学研究项目 2014FJ3016:湖南省科技厅项目

本校为第一机构

目的：建立体外胶原酶消化法分离培养大鼠肺动脉平滑肌细胞( PASMCs)及免疫鉴定的方法。方法采用I型胶原酶对PASMCs进行体外培养。在无菌环境下,分离提取雄性Sprague Dawley( SD)大鼠肺动脉主干,剥离外膜、剔除内皮细胞,经酶消化法,体外培养PASMCs。倒置相差显微镜观察PASMCs生长状态及特点；台盼兰测定细胞活力；免疫细胞染色法进行平滑肌α-肌动蛋白(α-SM actin)鉴定。结果酶消化法分离培养的PASMCs 3 d后,可见细胞贴壁呈梭形生长,6 d后生长迅速呈典型的峰-谷状生长,9 d后达90‰融合。通过形态学观察及免疫荧光染色法鉴定表明：培养的细胞为PASMCs；细胞存活率为98.5‰；原代培养8~10 d后即可...

Objective To set up the methods of rat pulmonary artery smooth muscle cells ( PASMCs) isolation,culture and immunological identification in vitro. Methods The PASMCs cultured in vitro with type I collagenase. Be-fore the PASMCs cultured,male SD rat pulmonary trunk separated were subjected to outer membrane peel and endothelial cells remove by enzymatic digestion in a sterile environment. We observed the status and characteristics of PASMCs with inverte phase contrast microscope,determined the cell viability with trypan blue staining,and indenti...