| Description |
Heart disease remains the leading cause of death in the United States, and such disease is caused by electrical, chemical, or biomechanical imbalances in the heart. The two most abundant cells comprising the heart that are responsible for these functions are cardiomyocytes and cardiac fibroblasts. In normal cardiac tissue, these fibroblasts are found surrounding the myocytes and bridging gaps between myocyte layers in such a way that each cardiomyocyte is closely related to one or more neighboring fibroblasts. Unsurprisingly, defects in cardiac fibroblast function have been implicated in a variety of heart disease conditions including fibrosis, aberrant electrical signaling, and hypertrophy. However, it is difficult to study cardiac fibroblasts due to limited availability of primary human cells and challenges to propagate them long-term in vitro. The aim of this study was to develop a new protocol for the differentiation of human induced-pluripotent stem cells (iPSCs) to cardiac fibroblasts that is more efficient and reproducible than current methods. The proposed method requires 10 days (compared to the established Zhang et. al. protocol which takes 20 days) and uses far less expensive reagents. The new proposed protocol resulted in successful differentiation of iPSCs to cardiac fibroblasts and was verified with flow cytometry using iPSC and cardiac fibroblast markers. Having an effective protocol for the differentiation of cardiac fibroblasts allows for the future in vitro study of cardiac fibroblast activation to myofibroblasts that is implicated in wide variety of heart disease. Understanding the interactions of cardiac fibroblasts with cardiomyocytes provides insights into the causes of cardiac dysfunction as well as a possible direction for the treatment of patients. |
