High-efficiency RNA-based reprogramming of human primary fibroblasts

Nature Communications
Igor KogutGanna Bilousova

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.

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Citations

Jul 29, 2018·Stem Cell Research & Therapy·Haissam Abou-SalehAli H Eid
Oct 28, 2019·The British Journal of Dermatology·T FunkE A O'Toole
Oct 5, 2018·Journal of Translational Medicine·Neil F BoxFabian V Filipp
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Feb 28, 2019·Nature Protocols·Onelia GaglianoNicola Elvassore
Dec 11, 2019·Proceedings of the National Academy of Sciences of the United States of America·Joanna JackówAngela M Christiano
Apr 8, 2020·Stem Cells International·Xinchao HuYuming Xu
Aug 25, 2018·Stem Cell Research & Therapy·Aline Vitrac, Isabelle Cloëz-Tayarani
Mar 21, 2019·Stem Cells Translational Medicine·Zohreh Sadat Badieyan, Todd Evans
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Oct 24, 2020·Expert Opinion on Emerging Drugs·Matthias TiteuxAlain Hovnanian
Nov 5, 2020·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Hanieh MoradianManfred Gossen
Nov 25, 2020·Stem Cell Reviews and Reports·Xiangyu ShiJiesi Luo
Feb 24, 2021·Disease Models & Mechanisms·Grant F MarshallCatherine M Abbott
Apr 8, 2021·GeroScience·Lucas Paulo de Lima Camillo, Robert B A Quinlan
May 21, 2021·Carcinogenesis·James E Trosko
Jul 25, 2021·International Journal of Molecular Sciences·Cameron L McKnightAnn E Frazier
Aug 8, 2021·International Journal of Molecular Sciences·Aline Yen Ling Wang
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Aug 1, 2021·The Journal of Allergy and Clinical Immunology·Stephan A RamosHolger A Russ
Dec 22, 2021·Nanotechnology·Yonger XueYizhou Dong

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Datasets Mentioned

BETA
GSE97265

Methods Mentioned

BETA
transfection
flow cytometry
transfections
RNA-Seq
Assay
biopsy
in vitro transcription
PCR
biopsies
fluorescence-activated cell sorting

Software Mentioned

Nanostring
FlowJo
Nanostring nCounter Gene
Nanostring nCounter
GSNAP
nSolver®
custom Perl script
Adobe Photoshop
Slidebook
R

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