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Mol Syst Biol. 2014 Nov 17;10:760. doi: 10.15252/msb.20145508.

Rapid neurogenesis through transcriptional activation in human stem cells.

Author information

1
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA.
2
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA Department of Biology, Brigham Young University, Provo, UT, USA Department of Pediatrics, University of California, San Diego, CA, USA.
3
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
4
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
5
Broad Institute of MIT and Harvard Cambridge Center, Cambridge, MA, USA McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
6
Department of Cell Biology, Harvard Medical School, Boston, MA, USA Division of Newborn Medicine, Boston Children's Hospital, Boston, MA, USA.
7
Department of Bioengineering, University of California, San Diego, CA, USA.
8
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland Swiss Institute of Bioinformatics, Basel, Switzerland University of Basel, Basel, Switzerland.
9
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA gchurch@genetics.med.harvard.edu.

Abstract

Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human-induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems-level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types.

KEYWORDS:

gene regulatory networks; microRNAs; neurogenesis; stem cell differentiation; transcriptomics

PMID:
25403753
PMCID:
PMC4299601
[Indexed for MEDLINE]
Free PMC Article

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