Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells / 中国应用生理学杂志

ABSTRACT

<p><b>OBJECTIVE</b>Set up a method to isolate and identify the small pulmonary arterial smooth muscle cells (PASMCs) in vitro.</p><p><b>METHODS</b>In sterile conditions, separated the male SD rat pulmonary artery, digested by collagenase I and cultured primary PASMCs. Measured cell viability; observed by phase contrast microscope; identified by immunocytochemistry and immunofluorescence staining as a label for smooth muscle alpha-actin.</p><p><b>RESULTS</b>PASMCs were identified by morphology and immunocytochemistry, immunofluorescence staining, with the cell viability is over 96.5%. The primary culture could be subcultured after 4-7 days and successfully passaged without change in morphology and growth characteristic.</p><p><b>CONCLUSION</b>This technique has advantage of the method is simple, short cultivate, good reproducibility, the primary cultured PASMCs quantity and the rapid growth.</p>