Abstract
Isolation and cultivation of primary aortic smooth muscle cells (SMCs) provide a valuable approach for investigating cellular mechanisms underlying vascular diseases, including aortic aneurysms. The present article describes a detailed and optimized protocol for the isolation and cultivation of murine aortic SMCs from thoracic and abdominal aortic segments. The current procedure includes careful aortic dissection, removal of surrounding tissue, and a two-step enzymatic digestion using collagenase, elastase, and soybean trypsin inhibitor, followed by mechanical dissociation and cell expansion under defined culture conditions. Particular emphasis is given on steps that improve cell yield, purity, and reproducibility, including adventitial removal and controlled digestion timing. Using this approach, the primary SMCs can be successfully isolated and expanded, exhibiting characteristic morphology and growth patterns. This protocol provides a reproducible and adaptable approach for generating primary aortic SMCs and serves as a platform for in vitro studies relevant to vascular remodeling and aortic aneurysm biology.