摘要
目的:建立一种重复性好、培养周期短及传代次数多的大鼠细小肺动脉平滑肌细胞(PASMCs)培养方法。方法:在无菌条件下,分离雄性SD大鼠肺细小动脉,剥离外膜和剔除内皮细胞,经胶原酶I消化,培养PASMCs。0.4%台盼蓝染色测定细胞活力;倒置相差显微镜观察;免疫细胞化学法和免疫荧光染色法,进行平滑肌α-肌动蛋白(α-SMactin)鉴定。结果:形态学观察、免疫细胞化学法及免疫荧光染色法鉴定表明培养细胞为PASMCs;细胞存活率在96.5%以上;原代培养后4～7d即可传代,并且生长特点、细胞形态不易发生改变。结论:采用胶原酶I消化法培养PASMCs,方法简单、酶消化时间易控制、培养周期短、重复性好,培养的原代PASMCs具有数量多和生长迅速的特点。
Objective：Set up a method to isolate and identify the small pulmonary arterial smooth muscle cells（PASMCs） in vitro.Methods：In sterile conditions,separated the male SD rat pulmonary artery,digested by collagenase I and cultured primary PASMCs.Measured cell viability;observed by phase contrast microscope;identified by immunocytochemistry and immunofluorescence staining as a label for smooth muscle α-actin.Results：PASMCs were identified by morphology and immunocytochemistry,immunofluorescence staining,with the cell viability is over 96.5%.The primary culture could be subcultured after 4～7 days and successfully passaged without change in morphology and growth characteristic.Conclusion：This technique has advantage of the method is simple,short cultivate,good reproducibility,the primary cultured PASMCs quantity and the rapid growth.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2010年第1期125-128,共4页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(30670933)
浙江省研究生创新科研项目(YK2008079)
关键词
肺动脉平滑肌细胞
原代培养
鉴定
大鼠
pulmonary arterial smooth muscle cells
primary cell culture
identification
rat
