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Embryonic stem cells (ESCs) are an important source of cardiomyocytes for regenerating injured myocardium. The successful use of ESC‐derived cardiomyocytes in cardiac tissue engineering requires an understanding of the important scaffold properties and culture conditions to promote cell attachment, differentiation, organization, and contractile function. The goal of this work was to investigate how scaffold architecture and coculture with fibroblasts influences the differentiated phenotype of murine ESC‐derived cardiomyocytes (mESCDCs). Electrospinning was used to process an elastomeric biodegradable polyurethane (PU) into aligned or unaligned fibrous scaffolds. Bioreactor produced mESCDCs were seeded onto the PU scaffolds either on their own or after pre‐seeding the scaffolds with mouse embryonic fibroblasts (MEFs). Viable mESCDCs attached to the PU scaffolds and were functionally contractile in all conditions tested. Importantly, the aligned scaffolds led to the anisotropic organization of rod‐shaped cells, improved sarcomere organization, and increased mESCDC aspect ratio (length‐to‐diameter ratio) when compared to cells on the unaligned scaffolds. In addition, pre‐seeding the scaffolds with MEFs improved mESCDC sarcomere formation compared to mESCDCs cultured alone. These results suggest that both fiber alignment and pre‐treatment of scaffolds with fibroblasts improve the differentiation of mESCDCs and are important parameters for developing engineered myocardial tissue constructs using ESC‐derived cardiac cells. Biotechnol. Bioeng. 2012; 109:813–822. © 2011 Wiley Periodicals, Inc.

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Fiber alignment and coculture with fibroblasts improves the differentiated phenotype of murine embryonic stem cell‐derived cardiomyocytes for cardiac tissue engineering

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