Lu Shuyang, Sun Xiaoning, Hong Tao, Song Kai, Yang Shouguo, Wang Chunsheng
Shanghai Institute of Cardiovascular Disease, Zhongshan Hospital, Fudan University, Fenglin Road 180, Xujiahui District, Shanghai 200032, China.
J Cardiothorac Surg. 2013 Apr 12;8:83. doi: 10.1186/1749-8090-8-83.
Acute type A aortic dissection (TAAD) is a life-threatening vascular disease. Smooth muscle cells (SMCs) are the main composition of aortic media and dysfunction of SMCs may lead to acute TAAD. The aim of this work was to investigate whether the SMCs of acute TAAD could be isolated and cultured for further research.
TAAD tissues were obtained from acute TAAD patients who underwent emergent surgical treatment. A simple and economical technique of collagenase digestion method was used to isolate and culture human SMCs. Confocal laser scanning microscopy was applied to identify SMC phenotypes. Purity of isolated and cultured SMCs was analyzed with flow cytometry and fluorescence microscopy respectively.
The purity of isolated SMCs was 78.2%, including α-smooth muscle cell actin positive 13.9%, calponin positive 35.0% and double positive 29.3%. For cultured SMCs, abundant expression of α-smooth muscle cell actin was observed universally under fluorescence microscope. Confocal laser scanning microscope testified that cultured cells were double positive of α-smooth muscle actin and calponin.
This is the first report of successful culture of SMCs isolated from human acute TAAD tissues. Living human SMCs of acute TAAD provides us with a new method for studying formation of acute TAAD.
急性A型主动脉夹层（TAAD）是一种危及生命的血管疾病。平滑肌细胞（SMC）是主动脉中膜的主要成分，SMC功能障碍可能导致急性TAAD。本研究旨在探讨能否从急性TAAD患者中分离培养SMC用于进一步研究。
从接受急诊手术治疗的急性TAAD患者获取TAAD组织。采用一种简单经济的胶原酶消化法技术分离培养人SMC。应用共聚焦激光扫描显微镜鉴定SMC表型。分别用流式细胞术和荧光显微镜分析分离培养的SMC纯度。
分离的SMC纯度为78.2%，其中α-平滑肌肌动蛋白阳性占13.9%，钙调蛋白阳性占35.0%，双阳性占29.3%。对于培养的SMC，在荧光显微镜下普遍观察到α-平滑肌肌动蛋白表达丰富。共聚焦激光扫描显微镜证实培养的细胞为α-平滑肌肌动蛋白和钙调蛋白双阳性。
这是首次成功培养从人急性TAAD组织中分离的SMC的报道。急性TAAD活的人SMC为我们研究急性TAAD的形成提供了一种新方法。
Zotero 插件
