1. Small molecules facilitate rapid and synchronous iPSC generation.
- Author
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Bar-Nur O, Brumbaugh J, Verheul C, Apostolou E, Pruteanu-Malinici I, Walsh RM, Ramaswamy S, and Hochedlinger K
- Subjects
- Animals, Antioxidants pharmacology, Apoptosis, Cell Cycle Checkpoints, Cell Differentiation genetics, Cell Proliferation, Cells, Cultured, Glycogen Synthase Kinase 3 antagonists & inhibitors, Glycogen Synthase Kinase 3 beta, Green Fluorescent Proteins genetics, Kruppel-Like Factor 4, Kruppel-Like Transcription Factors biosynthesis, Kruppel-Like Transcription Factors genetics, Mice, Octamer Transcription Factor-3 biosynthesis, Octamer Transcription Factor-3 genetics, Proto-Oncogene Proteins c-myc biosynthesis, Proto-Oncogene Proteins c-myc genetics, SOXB1 Transcription Factors biosynthesis, SOXB1 Transcription Factors genetics, Ascorbic Acid pharmacology, Cellular Reprogramming genetics, Embryonic Stem Cells cytology, Induced Pluripotent Stem Cells cytology, Pyridines pharmacology, Pyrimidines pharmacology
- Abstract
The reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) upon overexpression of OCT4, KLF4, SOX2 and c-MYC (OKSM) provides a powerful system to interrogate basic mechanisms of cell fate change. However, iPSC formation with standard methods is typically protracted and inefficient, resulting in heterogeneous cell populations. We show that exposure of OKSM-expressing cells to both ascorbic acid and a GSK3-β inhibitor (AGi) facilitates more synchronous and rapid iPSC formation from several mouse cell types. AGi treatment restored the ability of refractory cell populations to yield iPSC colonies, and it attenuated the activation of developmental regulators commonly observed during the reprogramming process. Moreover, AGi supplementation gave rise to chimera-competent iPSCs after as little as 48 h of OKSM expression. Our results offer a simple modification to the reprogramming protocol, facilitating iPSC induction at unparalleled efficiencies and enabling dissection of the underlying mechanisms in more homogeneous cell populations.
- Published
- 2014
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