期刊
NATURE COMMUNICATIONS
卷 9, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-018-03190-3
关键词
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资金
- National Institutes of Health [R01AR059947, T32AR007411-33]
- US Department of Defense [W81XWH-12-1-0606]
- Foundation for Ichthyosis and Related Skin Types (F.I.R.S.T.)
- Dystrophic Epidermolysis Bullosa Research Association (DEBRA) International
- King Baudouin Foundation
- Epidermolysis Bullosa (EB) Research Partnership
- EB Medical Research Foundation
- SOHANA Research Fund
- Linda Crnic Institute for Down Syndrome
- Gates Frontiers Fund
- University of Colorado (UC) Skin Diseases Research Core Center [P30AR057212]
- Colorado's NIH/NCI Cancer Center Molecular Pathology Shared Resource [P30CA046934]
- UC Advanced Light Microscopy Core
- NIH/NCATS Colorado CTSI [UL1 TR001082]
- Biostatistics/Bioinformatics and Genomics and Microarray Shared Resources of Colorado's NIH/NCI Cancer Center [P30CA046934]
- National Institutes of Health [R01AR059947, T32AR007411-33]
- US Department of Defense [W81XWH-12-1-0606]
- Foundation for Ichthyosis and Related Skin Types (F.I.R.S.T.)
- Dystrophic Epidermolysis Bullosa Research Association (DEBRA) International
- King Baudouin Foundation
- Epidermolysis Bullosa (EB) Research Partnership
- EB Medical Research Foundation
- SOHANA Research Fund
- Linda Crnic Institute for Down Syndrome
- Gates Frontiers Fund
- University of Colorado (UC) Skin Diseases Research Core Center [P30AR057212]
- Colorado's NIH/NCI Cancer Center Molecular Pathology Shared Resource [P30CA046934]
- UC Advanced Light Microscopy Core
- NIH/NCATS Colorado CTSI [UL1 TR001082]
- Biostatistics/Bioinformatics and Genomics and Microarray Shared Resources of Colorado's NIH/NCI Cancer Center [P30CA046934]
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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