Cell Culture Models:Cardiac Fibroblasts (CFs): These cells are central to fibrosis. Isolate primary CFs from cardiac tissue or use commercially available cell lines. Induced Pluripotent Stem Cell-Derived Cardiac Fibroblasts (iPSC-CFs): iPSCs can be differentiated into CFs, providing a valuable tool for drug screening1. Coculture Systems: Combine CFs with other cardiac cell types (e.g., cardiomyocytes, endothelial cells) to mimic the cardiac microenvironment.
Stimulating Fibrosis:TGF-β1 (Transforming Growth Factor-β1): This cytokine induces fibrosis by promoting collagen synthesis and myofibroblast activation. Angiotensin II: Known to stimulate fibrosis pathways. Mechanical Stretch: Apply cyclic mechanical strain to mimic the heart’s dynamic environment.
Extracellular Matrix (ECM) Components:Cultivate CFs on ECM-coated surfaces (e.g., collagen, fibronectin). Use 3D hydrogels to better mimic the native ECM.
Assessing Fibrosis:Immunofluorescence Staining: Detect collagen (e.g., type I and III) and α-smooth muscle actin (α-SMA) expression. Quantitative PCR (qPCR): Measure fibrosis-related gene expression (e.g., COL1A1, COL3A1, α-SMA). ELISA: Quantify secreted fibrotic proteins (e.g., TGF-β1, CTGF).
High-Content Imaging (HCI) and Machine Learning (ML):Use HCI to capture complex features of fibrosis in vitro. ML-based image analysis enhances throughput and precision2.
3D Cardiac Organoids:Develop more physiological 3D models resembling cardiac tissue. Organoids allow better recapitulation of cell-cell interactions and ECM remodeling.
Drug Screening:Test antifibrotic compounds using in vitro models. Identify potential targets for therapeutic intervention.
Remember, cardiac fibrosis is multifaceted, involving cell signaling, ECM dynamics, and tissue mechanics. Combining relevant models and advanced imaging techniques will pave the way for novel treatments against cardiac fibrosis.