Modeling cardiac fibrosis in mice: (Myo)fibroblast phenotype after ischemia

Abstract

Cardiac (myo)fibroblasts play a key role in the regulation of wound healing and pathogenic remodeling after myocardial infarction. Impaired scar formation and alterations of the extracellular matrix network precipitate cardiac dysfunction leading to increased morbidity and mortality. Therapeutic approaches addressing (myo)fibroblast phenotype appear to be useful in preventing progressive structural, electrical, and functional impairment and heart failure. Permanent ligation of the left anterior descending coronary artery has proven to be a valuable experimental model to investigate the arrays of pathways/mechanisms involved in cardiac repair and extracellular matrix remodeling in ischemic heart failure. Here we describe the surgical procedure to occlude the left coronary artery in mice. Moreover, we present an accurate method to isolate (myo)fibroblasts from ischemic myocardium, with maintenance of the functional phenotype, using the specific marker for mouse cardiac fibroblasts mEF-SK4. The protocol can be completed within a few hours, and the isolated fibroblasts/myofibroblasts are suitable for downstream molecular biology applications, like gene expression profiling and cell culture.