Isolation and Validation of Vascular Smooth Muscle Cells

The WT and mdx mice were sacrificed by inhalation of carbon dioxide overdose, causing rapid unconsciousness and death [14 ]. VSMCs were isolated as previously described [4 (link)]. The isolated VSMCs were cultured in a humidified atmosphere (37 °C) and for eight–ten days after plating before experimentation. A VSMCs phenotype was confirmed by validating the expression of muscle myofilament proteins (α-smooth muscle actin and β-tropomyosin) using Western blotting [13 (link)]

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Uryash A., Mijares A., Estève E., Adams J.A, & Lopez J.R. (2023). Smooth Muscle Cells of Dystrophic (mdx) Mice Are More Susceptible to Hypoxia; The Protective Effect of Reducing Ca2+ Influx. Biomedicines, 11(2), 623.

Publication 2023

Actin Atmosphere Carbon dioxide Inhalation Mdx mice Muscle Muscle proteins Myofilament Myofilament proteins Overdose Phenotype Smooth muscle Tropomyosin

Corresponding Organization : Mount Sinai Medical Center

Other organizations : Instituto Venezolano de Investigaciones Científicas, Université Grenoble Alpes, Centre National de la Recherche Scientifique, Université de Montpellier, Inserm, Translational Innovation in Medicine and Complexity

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Variable analysis

independent variables

WT mice
Mdx mice

dependent variables

VSMC phenotype
Expression of muscle myofilament proteins (α-smooth muscle actin and β-tropomyosin)

control variables

VSMC culture conditions (humidified atmosphere, 37 °C)
VSMC culture duration (eight–ten days after plating)

controls

Positive control: Western blotting to validate the expression of muscle myofilament proteins (α-smooth muscle actin and β-tropomyosin) [13 (link)]

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