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作 者:刘幸平[1] 刘清[1] 郑燕珊[1] 黄展勤[1]
机构地区:[1]汕头大学医学院药理学教研室,广东汕头515041
出 处:《汕头大学医学院学报》2008年第3期137-139,F0002,共4页Journal of Shantou University Medical College
基 金:国家自然科学基金资助项目(30672465);广东省自然科学基金资助项目(07300864);广东省医学科研基金资助项目(B2007120);汕头市科技计划项目(2008年度)
摘 要:目的:改进大鼠血管平滑肌细胞(VSMCs)的培养方法。方法:采用机械刮除、差异贴壁等手段进行组织贴块法原代培养,胰蛋白酶消化传代,并应用相差显微镜和即用型SABC免疫组化染色试剂盒对细胞进行形态学和免疫组化鉴定。结果:镜下培养细胞呈典型的"谷-峰状"生长,免疫组化染色显示胞浆内-αactin阳性表达,锥虫蓝染色检测细胞传代成活率95%。结论:本法可获纯度高、结构和功能良好的VSMCs。Objective: To improve the culture method of vascular smooth muscle cells(VSMCs)in rats. Methods: The primary culture and subculture were done by modified tissue-piece inoculation and trypsin digestion respectively. The cells were purified by the mechanical treatment and differential attachment. The cultured ceils were identified by morphology and immunohistochemistry with phase contrast microscope and SABC kit. Results: The cultured cells possessed "peak and valley" characteristics for VSMCs under microscope. Immunohistochemical staining with antibody against SM- α-actin demonstrated these ceils were positive. The cell viability was 95% in subculture by trypan blue. Conclusion: The highly purified VSMCs with good structure and function are obtained by this method.
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