High-efficiency RNA-based reprogramming of human primary fibroblasts

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, Dennis R. Roop () and Ganna Bilousova ()
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Igor Kogut: University of Colorado School of Medicine
Sandra M. McCarthy: University of Colorado School of Medicine
Maryna Pavlova: University of Colorado School of Medicine
David P. Astling: University of Colorado School of Medicine
Xiaomi Chen: University of Colorado School of Medicine
Ana Jakimenko: 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
Ganna Bilousova: University of Colorado School of Medicine

Nature Communications, 2018, vol. 9, issue 1, 1-15

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-03190-3

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