Isolation and culture of smooth muscle cells from human acute type A aortic dissection

Abstract

BackgroundAcute 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.MethodsTAAD 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.ResultsThe 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.ConclusionsThis 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.

Isolation, culture and identification of pulmonary arterial smooth muscle cells from rat distal pulmonary arteries

“…The presence of smooth muscle a-actin is the most widely used criterion for identification of smooth muscle cells. We found that smooth muscle a-actin expression was strongly positive and cable-like myofilaments filled in our cultured cells, consistent with observations from other investigators (Dufresne and Warocquier-Clérout 2001;Lu et al 2013;Crosas-Molist et al 2015). However, the immunofluorescence of smooth muscle a-actin was negative in rat PAECs which were stained strongly for von Willebrand Factor.…”

Myosin Binding Protein-C Forms Amyloid-Like Aggregates In Vitro

“…Cytotoxicity was assessed on smooth muscle cells isolated from rat aorta as described in [ 76 ] using the crystal violet assay [ 77 ]. The cells were seeded into 96-well cell culture plates (Greiner, Pleidelsheim, Germany) at a density of 3000 cells per well.…”

Smooth muscle titin formsin vitroamyloid aggregates

“…SMT quality and any degradation after lyophilization were monitored using 7% SDS/PAGE [ 25 ]. Cytotoxicity was evaluated with smooth muscle cells isolated from bovine aorta as described in [ 29 ] using crystalline violet assay [ 30 ]. The cells were seeded into 96-well cell culture plates (Greiner) at density of 3000 cells per well.…”