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大鼠细小肺动脉平滑肌细胞原代培养和鉴定方法的研究 预览 被引量:20

Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells
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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.
作者 钱国清 王良兴 陈婵 黄晓颖 叶舒婷 QIAN Guo-qing,WANG Liang-xing,CHENG Chan,HUANG Xiao-ying,YEI Shu-ting(Department of Respiratory,The First Affiliated Hospital,Wenzhou Medical College,Wenzhou 325000,China)
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
作者简介 【通讯作者】Tel:(0577)88069313,E-mail:38805@163.com
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