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In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter
JoVE Journal
Medicine
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JoVE Journal Medicine
In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter
DOI:

05:04 min

February 22, 2022

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Chapters

  • 00:04Introduction
  • 00:53Isolation and Selection of Neonatal Myh6-Cre/Rosa26A-Flox-Stop-Flox-GCaMP3 Mice Hearts
  • 01:23Isolation of Neonatal Cardiac Fibroblasts (NCFs)
  • 02:43Reprogramming NCFs to iCMs with MGT Encoding Retrovirus Infection
  • 03:28Results: In Vitro Assessment of Cardiac Reprogramming
  • 04:20Conclusion

Summary

Automatic Translation

We describe here, the establishment and application of an Tg(Myh6-cre)1Jmk/J /Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze/J (referred to as αMHC-Cre/Rosa26A-Flox-Stop-Flox-GCaMP3 below) mouse reporter line for cardiac reprogramming assessment. Neonatal cardiac fibroblasts (NCFs) isolated from the mouse strain are converted into induced cardiomyocytes (iCMs), allowing for convenient and efficient evaluation of reprogramming efficiency and functional maturation of iCMs via calcium (Ca2+) flux.

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