Journal
/
Developmental Biology
/
Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes
JoVE Journal
Developmental Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Developmental Biology
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

•

09:16 min

•

June 03, 2018

•

, , , , ,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/57687?article_type=v

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects
check_url/57687?article_type=v

Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5
check_url/57687?article_type=v

Protocol for the Direct Conversion of Murine Embryonic Fibroblasts into Trophoblast Stem Cells
check_url/57687?article_type=v

Reprogramming Mouse Embryonic Fibroblasts with Transcription Factors to Induce a Hemogenic Program
check_url/57687?article_type=v

Stimulation of Notch Signaling in Mouse Osteoclast Precursors
check_url/57687?article_type=v

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts
check_url/57687?article_type=v

Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies
check_url/57687?article_type=v

Dissection and Culture of Mouse Embryonic Kidney
check_url/57687?article_type=v

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle
check_url/57687?article_type=v

Direct Lineage Reprogramming of Adult Mouse Fibroblast to Erythroid Progenitors

Read Article