中南医学科学杂志Issue(1)：78-81,4.DOI:10.15972/j.cnki.43-1509/r.2015.01.020
酶消化法分离培养原代肺动脉平滑肌细胞及免疫组化鉴定
Methods of Culture of Rat Pulmonary Artery Smooth Muscle Cells and Identification
摘要
Abstract
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 indentified theα-smooth muscle actin (α-SM actin) with immuncytochemistry staining. Results The cultured PASMCs subjected to enzymatic digestion and isolation presented shuttle shape at d 3,typical peak - valley -like growth at d6,and 90‰ confluence at d9. The morphological observation and immuncytochemistry staining identification showed that:the cells cultured were PASMCs;cell survival rate up to 98. 5‰;the primary cultures can be passaged at d8~ d10 and cells can be used for cell experiments at 3th generation to 10th for stable morphology and fast growth. Conclutions The method type I collage-nase digestion is easy operation and reliable,the primary cultured PASMCs presented fast growt and short cell cycle can be used for pulmonary arterial hypertension and pulmonary vascular remodeling study.关键词
酶消化法/大鼠肺动脉平滑肌细胞/免疫细胞化学/SD大鼠Key words
Enzyme Digestion/PASMCs/SD-rat/immunocytochemistry分类
医药卫生引用本文复制引用
王爱平,李严兵,谢巍,曹宇辉,彭田红,周小兵,龚邵新..酶消化法分离培养原代肺动脉平滑肌细胞及免疫组化鉴定[J].中南医学科学杂志,2015,(1):78-81,4.基金项目
湖南省教育厅科研项目(13C837),湖南省科技厅项目(2014FJ3016) （13C837）
