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High-efficiency RNA-based reprogramming of human primary fibroblasts

Author

Listed:
  • Igor Kogut

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • Sandra M. McCarthy

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • Maryna Pavlova

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • David P. Astling

    (University of Colorado School of Medicine)

  • Xiaomi Chen

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • Ana Jakimenko

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • Kenneth L. Jones

    (University of Colorado School of Medicine)

  • Andrew Getahun

    (University of Colorado School of Medicine)

  • John C. Cambier

    (University of Colorado School of Medicine)

  • Anna M. G. Pasmooij

    (University Medical Center)

  • Marcel F. Jonkman

    (University Medical Center)

  • Dennis R. Roop

    (University of Colorado School of Medicine
    University of Colorado School of Medicine)

  • Ganna Bilousova

    (University of Colorado School of Medicine
    University of Colorado School of Medicine
    University of Colorado School of Medicine)

Abstract

Induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine; however, their potential clinical application is hampered by the low efficiency of somatic cell reprogramming. Here, we show that the synergistic activity of synthetic modified mRNAs encoding reprogramming factors and miRNA-367/302s delivered as mature miRNA mimics greatly enhances the reprogramming of human primary fibroblasts into iPSCs. This synergistic activity is dependent upon an optimal RNA transfection regimen and culturing conditions tailored specifically to human primary fibroblasts. As a result, we can now generate up to 4,019 iPSC colonies from only 500 starting human primary neonatal fibroblasts and reprogram up to 90.7% of individually plated cells, producing multiple sister colonies. This methodology consistently generates clinically relevant, integration-free iPSCs from a variety of human patient’s fibroblasts under feeder-free conditions and can be applicable for the clinical translation of iPSCs and studying the biology of reprogramming.

Suggested Citation

  • Igor Kogut & Sandra M. McCarthy & Maryna Pavlova & David P. Astling & Xiaomi Chen & Ana Jakimenko & Kenneth L. Jones & Andrew Getahun & John C. Cambier & Anna M. G. Pasmooij & Marcel F. Jonkman & Denn, 2018. "High-efficiency RNA-based reprogramming of human primary fibroblasts," Nature Communications, Nature, vol. 9(1), pages 1-15, December.
  • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-03190-3
    DOI: 10.1038/s41467-018-03190-3
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    Cited by:

    1. Claire Vinel & Gabriel Rosser & Loredana Guglielmi & Myrianni Constantinou & Nicola Pomella & Xinyu Zhang & James R. Boot & Tania A. Jones & Thomas O. Millner & Anaelle A. Dumas & Vardhman Rakyan & Je, 2021. "Comparative epigenetic analysis of tumour initiating cells and syngeneic EPSC-derived neural stem cells in glioblastoma," Nature Communications, Nature, vol. 12(1), pages 1-20, December.

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