Aim: Human embryonic stem cells (hESCs) represent a novel cell source to treat diseases such as heart failure and for use in drug screening. In this study, we aim to promote efficient generation of cardiomyocytes from hESCs by combining the current optimal techniques of controlled growth of undifferentiated cells and specific induction for cardiac differentiation. We also aim to examine whether these methods are scalable and whether the differentiated cells can be cryopreserved. Methods & results: hESCs were maintained without conditioned medium or feeders and were sequentially treated with activin A and bone morphogenetic protein-4 in a serum-free medium. This led to differentiation into cell populations containing high percentages of cardiomyocytes. The differentiated cells expressed appropriate cardiomyocyte markers and maintained contractility in culture, and the majority of the cells displayed working chamber (atrial and ventricular) type electrophysiological properties. In addition, the cell growth and differentiation process was adaptable to large culture formats. Moreover, the cardiomyocytes survived following cryopreservation, and viable cardiac grafts were detected after transplantation of cryopreserved cells into rat hearts following myocardial infarctions. Conclusion: These results demonstrate that cardiomyocytes of high quality can be efficiently generated and cryopreserved using hESCs maintained in serum-free medium, a step forward towards the application of these cells to human clinical use or drug discovery.

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Vol. 6, No. 1

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Published online 22 December 2010

Published in print January 2011

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Acknowledgement

The authors thank K Spink for critical review of this manuscript. The authors would also like to thank JA Thomson (University of Wisconsin-Madison, WI, USA) for kindly providing the IMR90 clone-1 human iPSC line to MA Laflamme.

Financial & competing interests disclosure

C Xu, S Police, M Hassanipour, Y Li, Y Chen, C Priest, C O’Sullivan, J Yang, K Delavan-Boorsma, A Davies, J Lebkowski and JD Gold are employees and shareholders of Geron Corporation. MA Laflamme is a collaborator of Geron Corporation. MA Laflamme was supported by NIH grants R01-HL064387 and K08-HL80431. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

All animal studies were approved by the Geron Animal Care and Use Committee and were in accordance with federal guidelines.

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Efficient generation and cryopreservation of cardiomyocytes derived from human embryonic stem cells

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