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Isolation and Culture of Smooth Muscle Cells from Human Acute type A Aortic Dissection

Published: Tuesday, April 23, 2013
Last Updated: Tuesday, April 23, 2013
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The aim of this work was to investigate whether the SMCs of acute TAAD could be isolated and cultured for further research.

Background
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.

Methods
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.

Results
The purity of isolated SMCs was 78.2%, including alpha-smooth muscle cell actin positive 13.9%, calponin positive 35.0% and double positive 29.3%. For cultured SMCs, abundant expression of alpha-smooth muscle cell actin was observed universally under fluorescence microscope. Confocal laser scanning microscope testified that cultured cells were double positive of alpha-smooth muscle actin and calponin.

Conclusions
This is the first report of successful culture of SMCs isolated from human acute TAAD tissues. Living human SMC.s of acute TAAD provides us with a new method for studying formation of acute TAAD.

This article is published online in the Journal of Cardiothoracic Surgery and is free to access.


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