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发育生物学
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Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes
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发育生物学
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JoVE 杂志 发育生物学
Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes

Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes

DOI:
10.3791/57687-v

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09:16 min

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June 03, 2018

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, , , , ,
1Division of Cardiology, Department of Medicine,University of Colorado Anschutz Medical Campus, 2Department of Population Health Sciences, Medical College of Georgia,Augusta University

Chapters

  • 00:05Title
  • 00:46Production of Retrovirus and Transduction of MEFs
  • 05:01Immunostaining of Reprogrammed MEFs
  • 06:44Results: Over-expression of GHMT2m and Inhibition of TGF-β Signaling Increases the Reprogramming Efficiency of MEFs into Functional Cardiomyocytes
  • 08:34Conclusion

Summary

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Here we present a robust method to reprogram primary embryonic fibroblasts into functional cardiomyocytes through overexpression of GATA4, Hand2, Mef2c, Tbx5, miR-1, and miR-133 (GHMT2m) alongside inhibition of TGF-β signaling. Our protocol generates beating cardiomyocytes as early as 7 days post-transduction with up to 60% efficiency.

Tags

Keywords: Pro-fibrotic SignalingFactor-mediated ReprogrammingInduced CardiomyocytesCardiac ReprogrammingFibroblast To Cardiomyocyte ConversionCardiomyocyte GenerationChemic Heart DiseaseFibroblast TransfectionRetrovirus ProductionPlatinum E CellsGHMT2M CocktailMouse Embryonic FibroblastsCollagen Coating

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