Abstract
The reprogramming of somatic cells to pluripotency using defined transcription factors holds great promise for biomedicine. However, human reprogramming remains inefficient and relies either on the use of the potentially dangerous oncogenes KLF4 and CMYC or the genetic inhibition of the tumor suppressor gene p53. We hypothesized that inhibition of signal transduction pathways that promote differentiation of the target somatic cells during development might relieve the requirement for non-core pluripotency factors during induced pluripotent stem cell (iPSC) reprogramming. Here, we show that inhibition of Notch greatly improves the efficiency of iPSC generation from mouse and human keratinocytes by suppressing p21 in a p53-independent manner and thereby enriching for undifferentiated cells capable of long-term self-renewal. Pharmacological inhibition of Notch enabled routine production of human iPSCs without KLF4 and CMYC while leaving p53 activity intact. Thus, restricting the development of somatic cells by altering intercellular communication enables the production of safer human iPSCs.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cells, Cultured
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Cyclin-Dependent Kinase Inhibitor p21 / genetics
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Cyclin-Dependent Kinase Inhibitor p21 / metabolism
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Dipeptides / pharmacology
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Genes, myc
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Genes, p53
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Histone-Lysine N-Methyltransferase
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Humans
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Keratinocytes / drug effects
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors / genetics
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Kruppel-Like Transcription Factors / metabolism
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Methyltransferases / genetics
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Methyltransferases / metabolism
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Mice
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Oncogenes / physiology*
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Pluripotent Stem Cells / physiology*
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Receptors, Notch / antagonists & inhibitors*
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Signal Transduction / physiology
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Transcription Factors / genetics
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Transcription Factors / metabolism
Substances
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CDKN1A protein, human
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Cyclin-Dependent Kinase Inhibitor p21
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DNA-Binding Proteins
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Dipeptides
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KLF4 protein, human
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Klf4 protein, mouse
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors
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MAML1 protein, human
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N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester
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Receptors, Notch
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Transcription Factors
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DOT1L protein, human
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Methyltransferases
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Histone-Lysine N-Methyltransferase