title: Isolation and culture of smooth muscle cells from human acute type A aortic dissection.
abstract: 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 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Shanghai Institute of Cardiovascular Disease, Zhongshan Hospital, Fudan University, Fenglin Road 180, Xujiahui District, Shanghai 200032, China.
authors.forename: Shuyang
authors.initials: S
authors.lastname: Lu
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Sun
authors.forename: Tao
authors.initials: T
authors.lastname: Hong
authors.forename: Kai
authors.initials: K
authors.lastname: Song
authors.forename: Shouguo
authors.initials: S
authors.lastname: Yang
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Wang
mesh_terms: D000199:Actins / Q000032:analysis / Q000737:chemistry; D000328:Adult; D000784:Aortic Dissection / Q000473:pathology*; D001011:Aorta / Q000166:cytology*; D001014:Aortic Aneurysm / Q000473:pathology*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000737:chemistry / Q000166:cytology*
chemical_list: D000199:Actins