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

In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter

DOI:
10.3791/62643-v

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05:04 min

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February 22, 2022

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, ,
1Department of Cardiac Surgery, Cardiovascular Center,The University of Michigan, Ann Arbor, 2Department of Cardiovascular Medicine, Xiangya Hospital,Central South University

Capitoli

  • 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

Traduzione automatica

Traduzione automatica

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.

Tags

Keywords: Cardiac ReprogrammingCalcium FluxGCaMP3 ReporterNeonatal Cardiac FibroblastsIsolationCell CultureFunctional MaturationCardiomyocyte DifferentiationCardiac Function

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