Observation of cardiomyoblast differentiation on different ECM microenvironments
- Observation of cardiomyoblast differentiation on different ECM microenvironments
- 수하에리; 반세영; 라메쉬; 두 핑; 김인걸; 박귀덕
- Extracellular matrix; fibroblast-derived matrix; cardiomyoblasts (H9c2); cardiomyogenic differentiation; matrix stiffness
- Issue Date
- Extracellular matrix (ECM) is a complex network of diverse microenvironments that provide specific chemical and physical cues. In this study, fibroblast-derived ECM (FDM), a model ECM has been utilized, against conventional culture platforms (gelatin and fibronectin) for cardiomyoblast (H9c2) culture and induction of cardiomyogenic differentiation. Morphological analysis using F-actin staining shows that cells in growth medium (GM) are more circular on FDM until day 3 but turn to a flat and elongated shape in differentiation medium (DM) at day 7. Cell proliferation assay exhibits that H9c2 proliferates well with time in GM whereas the cell numbers are rather constant in DM. Focal adhesion (FA) was also examined using immunofluorescence of vinculin: cells on gelatin have a larger FA area compared to FN or FDM in DM at early time point. When cardiomyogenic differentiation was evaluated using cardiomyocyte markers (α-actinin and cTnT) and specific gene expression (Actc1, Myl2, and Tnnt), FDM presented much higher differentiation efficiency and more upregulated gene expression, compared to that of gelatin and fibronectin. Additionally, FDM also presents higher percentage of cells that express gap junction protein, connexin 43. Further analysis employs a crosslinking agent in order to control matrix stiffness and examines optimized matrix stiffness for cardiomyogenic differentiation. In conclusion, cellular behavior of cardiomyoblast can be regulated by different contexts of ECM microenvironment. This work indicates that FDM is a promising resource over gelatin or FN for cardiomyoblast differentiation.
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