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RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells

RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells

DOI:
10.3791/58687-v

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11:38 min

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November 26, 2018

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, , ,
1Department of Dermatology,University of Colorado School of Medicine, Anschutz Medical Campus, 2Charles C. Gates Center for Regenerative Medicine,University of Colorado School of Medicine, Anschutz Medical Campus, 3Stem Cell Biobank and Disease Modeling Core,University of Colorado School of Medicine, Anschutz Medical Campus

Chapters

  • 00:04Title
  • 00:45Reagents Preparation
  • 02:01Plating Fibroblasts for Reprogramming: Day 0
  • 05:09Initiation of Reprogramming: Day 1
  • 09:28Results: Efficient Reprogramming of Human Primary Fibroblasts into iPSCs
  • 11:16Conclusion

Summary

Automatic Translation

Automatic Translation

Here we describe a clinically relevant, high-efficiency, feeder-free method to reprogram human primary fibroblasts into induced pluripotent stem cells using modified mRNAs encoding reprogramming factors and mature microRNA-367/302 mimics. Also included are methods to assess reprogramming efficiency, expand clonal iPSC colonies, and confirm expression of the pluripotency marker TRA-1-60.

Tags

Keywords: RNA-based ReprogrammingInduced Pluripotent Stem CellsIPSCsPrimary FibroblastsTransfection EfficiencyPH AdjustmentLaminin 521Fibroblast Expansion MediumPlating MediumFGFB18R

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