大鼠细小肺动脉平滑肌细胞原代培养和鉴定方法的研究

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。

Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells

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.