Abstract
Vascular smooth muscle cells (VSMCs) play important roles in cardiovascular disorders and biology. Outlined in this paper is a step-by-step procedure for isolating aortic VSMCs from adult C57BL6J male mice by enzymatic digestion of the aorta using collagenase. The plating, culturing, and subculturing of the isolated cells are discussed in detail along with techniques to characterize VSMC phenotype by gene expression and immunofluorescence. Traction force microscopy was used to characterize contractility of single subcultured VSMCs at baseline.
Publication types
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Guideline
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Actins / genetics
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Actins / metabolism
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Animals
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Aorta / cytology
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Aorta / metabolism
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Aorta / physiology
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Biomarkers / metabolism
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Cell Culture Techniques / standards*
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Cell Proliferation
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Cell Separation / methods
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Cell Separation / standards*
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Cells, Cultured
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Cytoskeletal Proteins / genetics
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Cytoskeletal Proteins / metabolism
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Gene Expression Regulation
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Male
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Mice, Inbred C57BL
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Microfilament Proteins / genetics
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Microfilament Proteins / metabolism
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Muscle Proteins / genetics
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Muscle Proteins / metabolism
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Muscle, Smooth, Vascular / cytology
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Muscle, Smooth, Vascular / metabolism
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Muscle, Smooth, Vascular / physiology*
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Myocytes, Smooth Muscle / metabolism
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Myocytes, Smooth Muscle / physiology*
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Phenotype
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Vasoconstriction
Substances
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Acta2 protein, mouse
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Actins
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Biomarkers
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Cytoskeletal Proteins
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Microfilament Proteins
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Muscle Proteins
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Smtn protein, mouse
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Tagln protein, mouse