Observation of cardiomyoblast differentiation on different ECM microenvironments

Title
Observation of cardiomyoblast differentiation on different ECM microenvironments
Authors
수하에리반세영라메쉬두 핑김인걸박귀덕
Keywords
Extracellular matrix; fibroblast-derived matrix; cardiomyoblasts (H9c2); cardiomyogenic differentiation; matrix stiffness
Issue Date
2014-09
Publisher
TERMIS-AP
Abstract
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.
URI
http://pubs.kist.re.kr/handle/201004/48468
Appears in Collections:
KIST Publication > Conference Paper
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