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Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling

A Corrigendum to this article was published on 01 May 2009

This article has been updated


Current neural induction protocols for human embryonic stem (hES) cells rely on embryoid body formation, stromal feeder co-culture or selective survival conditions. Each strategy has considerable drawbacks, such as poorly defined culture conditions, protracted differentiation and low yield. Here we report that the synergistic action of two inhibitors of SMAD signaling, Noggin and SB431542, is sufficient to induce rapid and complete neural conversion of >80% of hES cells under adherent culture conditions. Temporal fate analysis reveals the appearance of a transient FGF5+ epiblast-like stage followed by PAX6+ neural cells competent to form rosettes. Initial cell density determines the ratio of central nervous system and neural crest progeny. Directed differentiation of human induced pluripotent stem (hiPS) cells into midbrain dopamine and spinal motoneurons confirms the robustness and general applicability of the induction protocol. Noggin/SB431542-based neural induction should facilitate the use of hES and hiPS cells in regenerative medicine and disease modeling and obviate the need for protocols based on stromal feeders or embryoid bodies.

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Figure 1: Dual-SMAD inhibition allows for highly efficient feeder-free neural induction in adherent cultures in 7 d.
Figure 2: Neuralization of hES cells by dual-SMAD inhibition permits a pre-rosette, neural stem cell with dopaminergic and motoneuronal potential.
Figure 3: IPS cells can be differentiated to neural tissue using dual-SMAD inhibition and are patternable to dopaminergic neurons and motoneurons.
Figure 4: Model of proposed mechanisms that contribute to the action of Noggin and SB431542.

Change history

  • 16 March 2009

    In the version of this article initially published, the unit (nM) for the amount of TGF-b inhibitor (Tocris) reported in the Methods section “Neural induction” was incorrect. The correct unit is μM. The error has been corrected in the HTML and PDF versions of the article.


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We are grateful to F. Vaccarino for providing Otx2 antibody and E. Lai for BF1 antibody. This work was supported in part by the Starr Foundation, NINDS grant 1R01NS052671, the Starr Stem Scholar fellowship (S.M.C.) and the a New York Stem Cell Foundation fellowship (C.A.F.).

Author information




S.M.C. and L.S. designed the study. E.P.P., M.T., L.S. and M.S. designed and generated the hiPS clones. S.M.C. and L.S. analyzed the data and wrote the manuscript. S.M.C. and C.A.F. performed the experiments.

Corresponding authors

Correspondence to Stuart M Chambers or Lorenz Studer.

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Chambers, S., Fasano, C., Papapetrou, E. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27, 275–280 (2009).

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