Key result
Two-step enzymatic digestion protocol successfully isolates and expands primary murine aortic smooth muscle cells.
Why the study?
Isolating and cultivating primary aortic smooth muscle cells is valuable for investigating the cellular mechanisms of vascular diseases and aortic aneurysms, requiring an optimized protocol to improve cell yield, purity, and reproducibility.
Provides a reproducible laboratory protocol for isolating and culturing primary murine aortic smooth muscle cells for in vitro vascular remodeling and aortic aneurysm studies.
Answers are grounded in this paper and cite their sources.
May enable mechanistic studies of aortic aneurysms in mice; leaves open human translation and validation.
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.
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Brunner et al. (2026) studied Vascular remodeling and aortic aneurysm biology (murine model). Two-step enzymatic digestion (collagenase, elastase, soybean trypsin inhibitor) and mechanical dissociation protocol was evaluated on Isolation and expansion of primary murine aortic smooth muscle cells. A two-step enzymatic digestion protocol using collagenase, elastase, and soybean trypsin inhibitor successfully isolates and expands primary murine aortic smooth muscle cells.
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