Escape from Pluripotency via Inhibition of TGF-β/BMP and Activation of Wnt Signaling Accelerates Differentiation and Aging in hPSC Progeny Cells

Stem Cell Reports. 2017 Nov 14;9(5):1675-1691. doi: 10.1016/j.stemcr.2017.09.024. Epub 2017 Oct 26.


Human pluripotent stem cells (hPSCs) represent a potentially valuable cell source for applications in cell replacement therapy, drug development, and disease modeling. For all these uses, it is necessary to develop reproducible and robust protocols for differentiation into desired cell types. However, differentiation protocols remain unstable and inefficient, which makes minimizing the differentiation variance among hPSC lines and obtaining purified terminally differentiated cells extremely time consuming. Here, we report a simple treatment with three small molecules-SB431542, dorsomorphine, and CHIR99021-that enhanced hPSC differentiation into three germ layers with a chemically transitional embryoid-body-like state (CTraS). Induction of CTraS reduced the innate differentiation propensities of hPSCs (even unfavorably differentiated hPSCs) and shifted their differentiation into terminally differentiated cells, particularly neurons. In addition, CTraS induction accelerated in vitro pathological expression concurrently with neural maturation. Thus, CTraS can promote the latent potential of hPSCs for differentiation and potentially expand the utility and applicability of hPSCs.

Keywords: aging; differentiation; disease model; induced pluripotent stem cells; pluripotency; stem cell biotechnology; stem cell differentiation.

MeSH terms

  • Benzamides / pharmacology
  • Bone Morphogenetic Proteins / metabolism
  • Cell Differentiation*
  • Cell Line
  • Cells, Cultured
  • Cellular Senescence*
  • Dioxoles / pharmacology
  • Embryoid Bodies / cytology
  • Embryoid Bodies / drug effects
  • Embryoid Bodies / metabolism
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / drug effects
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • Neurons / cytology
  • Protein Kinase Inhibitors / pharmacology
  • Pyrazoles / pharmacology
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • Receptors, Transforming Growth Factor beta / antagonists & inhibitors*
  • Transforming Growth Factor beta / metabolism
  • Wnt Signaling Pathway*


  • 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide
  • Benzamides
  • Bone Morphogenetic Proteins
  • Chir 99021
  • Dioxoles
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyridines
  • Pyrimidines
  • Receptors, Transforming Growth Factor beta
  • Transforming Growth Factor beta
  • dorsomorphin