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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
引用本文:钱国清,王良兴,陈婵,黄晓颖,叶舒婷. 大鼠细小肺动脉平滑肌细胞原代培养和鉴定方法的研究[J]. 中国应用生理学杂志, 2010, 0(1): 125-128
作者姓名:钱国清  王良兴  陈婵  黄晓颖  叶舒婷
作者单位:浙江省温州医学院附属第一医院呼吸内科;
基金项目: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)
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。

关 键 词:肺动脉平滑肌细胞  原代培养  鉴定  大鼠

Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells
QIAN Guo-qing,WANG Liang-xing,CHENG Chan,HUANG Xiao-ying,YEI Shu-ting. Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells[J]. Chinese journal of applied physiology, 2010, 0(1): 125-128
Authors:QIAN Guo-qing  WANG Liang-xing  CHENG Chan  HUANG Xiao-ying  YEI Shu-ting
Affiliation: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)
Abstract: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.
Keywords:pulmonary arterial smooth muscle cells  primary cell culture  identification  rat  
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