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发育生物学
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Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts
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发育生物学
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JoVE 杂志 发育生物学
Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

DOI:
10.3791/55456-v

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09:29 min

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March 22, 2017

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1Department of Internal Medicine- Cardiology,UT Southwestern Medical Center

Chapters

  • 00:05Title
  • 00:40Retrovirus Production in Plat-E Cells
  • 02:04Hcn4-GFP Mouse Embryonic Fibroblast Seeding
  • 02:57Transduction and Reprogramming of Mouse Embryonic Fibroblasts
  • 04:30Immunostaining of Reprogrammed MEFs
  • 06:01Identification of Cardiac Subtypes Using Confocal Microscopy
  • 06:56Quantification of Sarcomere+ Cells
  • 07:32Results: Identification of Cardiomyocyte Sub-populations
  • 08:44Conclusion

Summary

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This manuscript describes a step-by-step protocol for the generation and quantification of diverse reprogrammed cardiac subtypes using a retrovirus-mediated delivery of Gata4, Hand2, Mef2c, and Tbx5.

Tags

Cardiomyocyte SubtypesTranscription Factor-mediated ReprogrammingMouse Embryonic FibroblastsSarcomere AssemblySubtype SpecificationLineage ConversionCardiac ReprogrammingGHMT CocktailHCN4GFP Mouse Embryonic FibroblastsImmunocytochemistryCollagenFibronectin

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