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Cardiomyoblast (h9c2) differentiation on tunable extracellular matrix microenvironment
Muhammad Suhaeri1,2, Ramesh Subbiah1,2, Se Young Van1,2
11Center for Biomaterials, Korea Institute of Science and Technology, Seoul, Republic of Korea.
Tissue Engineering. Part A
|April 4, 2015
Summary
Fibroblast-derived matrix (FDM) promotes cardiac cell differentiation more effectively than gelatin or fibronectin. Tunable matrix stiffness via crosslinking further enhances cardiomyogenesis for cardiac tissue engineering applications.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Extracellular matrices (ECM) are crucial for cell function, but their application in cardiac tissue engineering remains limited.
- Fibroblast-derived matrix (FDM) presents a novel ECM platform with unique properties.
- H9c2 cardiomyoblasts are a common cell line used in cardiac research.
Purpose of the Study:
- To investigate the cardiomyogenic potential of FDM compared to gelatin and fibronectin.
- To characterize the physical and biochemical properties of FDM.
- To explore the effect of tunable matrix stiffness on cardiomyogenesis.
Main Methods:
- Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) for FDM characterization.
- Immunofluorescence staining for ECM components (fibronectin, collagen, laminin).
- Cell culture of H9c2 cardiomyoblasts on FDM, gelatin, and fibronectin, followed by differentiation induction and marker analysis (α-actinin, cTnT, Myl2, Tnnt, connexin 43).
- Genipin crosslinking to tune FDM stiffness and subsequent analysis of cell response and differentiation.
Main Results:
- FDM exhibits unique surface texture and biomechanical properties, containing fibronectin, collagen, and laminin.
- H9c2 cells on FDM showed enhanced differentiation into cardiomyocytes, evidenced by increased expression of cardiac markers (α-actinin, cTnT, Myl2, Tnnt, connexin 43) compared to gelatin and fibronectin.
- Crosslinked FDM (X-FDM) with increased stiffness significantly promoted cardiomyoblast differentiation more than natural FDM.
- Integrin subunit α5 expression was upregulated on natural FDM compared to X-FDM, while β1 expression remained unchanged.
Conclusions:
- FDM serves as a favorable extracellular matrix microenvironment for cardiomyogenesis of H9c2 cells.
- Tunable mechanical compliance of FDM, achieved through crosslinking, plays a significant role in enhancing cardiomyogenesis.
- This study provides valuable insights into optimizing ECM properties for cardiac tissue engineering.
