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Updated: Jul 17, 2025
Isolation of Murine Coronary Vascular Smooth Muscle Cells
Published on: May 30, 2016
Isolation and Cultivation of Vascular Smooth Muscle Cells from the Mouse Circle of Willis
Wei Chang1, Yajuan Li1, Fengzhou Liu1,2
1Center for Aerospace Clinical Medicine, Department of Aerospace Medicine, Air Force Medical University, Xi'an, China.
Introduction:
Culturing cerebrovascular smooth muscle cells (CVSMCs) in vitro can provide a model for studying many cerebrovascular diseases. This study describes a convenient and efficient method to obtain mouse CVSMCs by enzyme digestion.
Methods:
Mouse circle of Willis was isolated, digested, and cultured with platelet-derived growth factor-BB (PDGF-BB) to promote CVSMC growth, and CVSMCs were identified by morphology, immunofluorescence analysis, and flow cytometry. The effect of PDGF-BB on vascular smooth muscle cell (VSMC) proliferation was evaluated by cell counting kit (CCK)-8 assay, morphological observations, Western blotting, and flow cytometry.
Results:
CVSMCs cultured in a PDGF-BB-free culture medium had a typical peak-to-valley growth pattern after approximately 14 days. Immunofluorescence staining and flow cytometry detected strong positive expression of the cell type-specific markers alpha-smooth muscle actin (α-SMA), smooth muscle myosin heavy chain 11 (SMMHC), smooth muscle protein 22 (SM22), calponin, and desmin. In the CCK-8 assay and Western blotting, cells incubated with PDGF-BB had significantly enhanced proliferation compared to those without PDGF-BB.
Conclusion:
We obtained highly purified VSMCs from the mouse circle of Willis using simple methods, providing experimental materials for studying the pathogenesis and treatment of neurovascular diseases in vitro. Moreover, the experimental efficiency improved with PDGF-BB, shortening the cell cultivation period.
Insights
This study presents an efficient method for culturing mouse cerebrovascular smooth muscle cells (CVSMCs) using enzyme digestion. Platelet-derived growth factor-BB (PDGF-BB) significantly enhances CVSMC proliferation, shortening cultivation time for research.
Area of Science:
- Neuroscience
- Cell Biology
- Vascular Biology
Background:
- Cerebrovascular smooth muscle cells (CVSMCs) are crucial for studying cerebrovascular diseases.
- In vitro models are essential for understanding disease pathogenesis and developing treatments.
Purpose of the Study:
- To establish a convenient and efficient method for obtaining and culturing mouse CVSMCs.
- To investigate the effect of platelet-derived growth factor-BB (PDGF-BB) on CVSMC proliferation.
Main Methods:
- Isolation and enzymatic digestion of the mouse circle of Willis.
- Culture of cells with and without PDGF-BB.
- Identification of CVSMCs using morphology, immunofluorescence (α-SMA, SMMHC, SM22, calponin, desmin), and flow cytometry.
- Assessment of cell proliferation via CCK-8 assay, Western blotting, and flow cytometry.
Main Results:
- CVSMCs exhibited a typical growth pattern within 14 days in culture.
- Immunofluorescence confirmed high expression of VSMC-specific markers.
- PDGF-BB significantly enhanced VSMC proliferation compared to controls.
Conclusions:
- A simple and efficient method for isolating and culturing high-purity mouse CVSMCs from the circle of Willis was developed.
- This method provides valuable in vitro models for neurovascular disease research.
- PDGF-BB accelerates cell cultivation, improving experimental efficiency.
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