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Biologia do Desenvolvimento
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Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5
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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5

Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5

DOI:
10.3791/53426-v

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10:05 min

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November 13, 2015

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, , , , ,
1Department of Pathology and Laboratory Medicine, McAllister Heart Institute,University of North Carolina, Chapel Hill

Capítulos

  • 00:05Título
  • 00:57Generation of Neonatal Mouse Cardiac Fibroblasts via Explant Culture
  • 03:20Generation of Neonatal Mouse Cardiac Fibroblasts via Enzymatic Digestion
  • 05:02Isolation of Thy1.2+ Fibroblasts by Magnetic-activated Cell Sorting (MACS)
  • 07:17Retroviral Mediated Reprogramming of Isolated Cardiac Fibroblasts
  • 08:52Results: Retroviral Reprogramming of Cardiac Fibroblasts to Induced Cardiac Myocytes
  • 09:38Conclusion

Summary

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We describe here a protocol for the generation of iCMs using retrovirus-mediated delivery of Gata4, Tbx5 and Mef2c in a polycistronic construct. This protocol yields a relatively homogeneous population of reprogrammed cells with improved efficiency and quality and is valuable for future studies of iCM reprogramming.

Tags

Keywords: Cardiomyocyte ReprogrammingInduced CardiomyocytesCardiac FibroblastsGata4Mef2cTbx5Polycistronic VectorReprogramming EfficiencyRegenerative Medicine

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