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Conversion of mouse fibroblasts into cardiomyocytes using a direct reprogramming strategy

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Abstract

Here we show that conventional reprogramming towards pluripotency through overexpression of Oct4, Sox2, Klf4 and c-Myc can be shortcut and directed towards cardiogenesis in a fast and efficient manner. With as little as 4 days of transgenic expression of these factors, mouse embryonic fibroblasts (MEFs) can be directly reprogrammed to spontaneously contracting patches of differentiated cardiomyocytes over a period of 11–12 days. Several lines of evidence suggest that a pluripotent intermediate is not involved. Our method represents a unique strategy that allows a transient, plastic developmental state established early in reprogramming to effectively function as a cellular transdifferentiation platform, the use of which could extend beyond cardiogenesis. Our study has potentially wide-ranging implications for induced pluripotent stem cell (iPSC)-factor-based reprogramming and broadens the existing paradigm.

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Figure 1: Development of robust conditions for direct reprogramming of fibroblasts to a mature cardiac fate.
Figure 2: Gauging reprogramming efficiency and success by incidence of beating and marker expression.
Figure 3: Calcium flux and electrophysiological characterization of contracting cardiomyocytes.
Figure 4: Development of pluripotency is detrimental to cardiogenesis.
Figure 5: Direct cardiac reprogramming represents a parallel process that occurs in the absence of iPSC generation.
Figure 6: A model for direct reprogramming to alternative fates.

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Change history

  • 11 February 2011

    In the version of this article initially published online, the commerically available cardiomyocytes Cor.At were incorrectly referred to as CorAT in Figure 4. Similarly, in the methods Axiogenesis was not correctly referenced as the manufacturers of Cor.At cells. This error has been corrected in both the HTML and PDF versions of the article.

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Acknowledgements

We thank C. Desponts, X. Yuan, S. Zhu, R. Ambasudhan, R. Abujarour and W. Li for discussions, technical assistance, and critical reading of the manuscript. We also thank H. Schöler for providing TTFs, and D. Watry for assistance with FACS analyses. S.D. is supported by funding from NICHD, NHLBI and NIMH/NIH, California Institute for Regenerative Medicine, Prostate Cancer Foundation, Fate Therapeutics, Esther B. O'Keeffe Foundation and the Scripps Research Institute. J.A.E. is a Lowe Family Foundation fellow.

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J.A.E. and S.D. conceived the project and wrote the manuscript. J.A.E. designed and carried out experiments. S.H., J.K., H.Z., K.O., G.W. and J.C. provided materials and assisted with experiments.

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Correspondence to Sheng Ding.

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The authors declare no competing financial interests.

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Efe, J., Hilcove, S., Kim, J. et al. Conversion of mouse fibroblasts into cardiomyocytes using a direct reprogramming strategy. Nat Cell Biol 13, 215–222 (2011). https://doi.org/10.1038/ncb2164

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