Abstract
One of the major obstacles in generating induced pluripotent stem (iPS) cells suitable for therapeutic application is the low efficiency of the process and the long time required, with many iPS lines acquiring genomic aberrations. In this chapter we describe a highly efficient iPS reprogramming system based on the transient expression in pre-B cells of the transcription factor C/EBPα, followed by the induction of the four Yamanaka factors (OSKM). In addition, the process is very rapid, yielding Oct4 positive cells within 2 days and Nanog-positive iPS cell colonies within a week.
Keywords:
B lymphocytes; Cell reprogramming; Deterministic process; Embryonic stem cells; Induced pluripotent stem cells; Klf4; Oct4; Sox2; Transcription factor C/EBPα; c-Myc.
MeSH terms
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Animals
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Antigens, Differentiation / analysis
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CCAAT-Enhancer-Binding Proteins / biosynthesis
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CCAAT-Enhancer-Binding Proteins / genetics
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Cell Culture Techniques / methods
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Cell Differentiation
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Cells, Cultured
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Cellular Reprogramming
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Cellular Reprogramming Techniques / methods
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Culture Media
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Doxycycline / pharmacology
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Fibroblasts / cytology
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Gene Expression / drug effects
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Genes, myc
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Induced Pluripotent Stem Cells / cytology*
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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 / physiology
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Mice
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / physiology
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Precursor Cells, B-Lymphoid / cytology*
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Proto-Oncogene Proteins c-myc / physiology
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SOXB1 Transcription Factors / genetics
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SOXB1 Transcription Factors / physiology
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Stromal Cells / cytology
Substances
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Antigens, Differentiation
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CCAAT-Enhancer-Binding Proteins
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CEBPA protein, mouse
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Culture Media
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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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Myc protein, mouse
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Octamer Transcription Factor-3
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Pou5f1 protein, mouse
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Proto-Oncogene Proteins c-myc
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SOXB1 Transcription Factors
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Sox2 protein, mouse
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Doxycycline