Journal
NATURE METHODS
Volume 11, Issue 11, Pages 1170-1176Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.3142
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Funding
- Gruss-Lipper postdoctoral fellowship
- US National Institutes of Health (NIH) [R01HD058013]
- Howard Hughes Medical Institute
- MGH Executive Committee on Research
- NIH [1F32HD078029-01A1]
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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-beta 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.
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