Embryonic Stem Cells/Induced Pluripotent Stem Cells
Version of Record online: 20 AUG 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 9, pages 1875–1884, September 2012
How to Cite
Surmacz, B., Fox, H., Gutteridge, A., Lubitz, S. and Whiting, P. (2012), Directing Differentiation of Human Embryonic Stem Cells Toward Anterior Neural Ectoderm Using Small Molecules. STEM CELLS, 30: 1875–1884. doi: 10.1002/stem.1166
Author contributions: B.S.: conception and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; H.F.: collection and assembly of data and data analysis and interpretation; A.G.: data analysis and interpretation and manuscript writing; S.L.: conception and design and data analysis and interpretation; P.W.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS July 3, 2012.
- Issue online: 20 AUG 2012
- Version of Record online: 20 AUG 2012
- Accepted manuscript online: 3 JUL 2012 08:01AM EST
- Manuscript Accepted: 5 JUN 2012
- Manuscript Received: 13 FEB 2012
Vol. 31, Issue 7, 1446, Version of Record online: 6 JUL 2013
- Embryonic stem cells;
- Neural induction;
- Gene expression
Based on knowledge of early embryo development, where anterior neural ectoderm (ANE) development is regulated by native inhibitors of bone morphogenic protein (BMP) and Nodal/Activin signaling, most published protocols of human embryonic stem cell differentiation to ANE have demonstrated a crucial role for Smad signaling in neural induction. The drawbacks of such protocols include the use of an embryoid body culture step and use of polypeptide secreted factors that are both expensive and, when considering clinical applications, have significant challenges in terms of good manufacturing practices compliancy. The use of small molecules to direct differentiation of pluripotent stem cells toward a specified lineage represents a powerful approach to generate specific cell types for further understanding of biological function, for understanding disease processes, for use in drug discovery, and finally for use in regenerative medicine. We therefore aimed to find controlled and reproducible animal-component-free differentiation conditions that would use only small molecules. Here, we demonstrate that pluripotent stem cells can be reproducibly and efficiently differentiated to PAX6+ (a marker of neuroectoderm) and OCT4− (a marker of pluripotent stem cells) cells with the use of potent small inhibitors of the BMP and Activin/Nodal pathways, and in animal-component-free conditions, replacing the frequently used Noggin and SB431542. We also show by transcript analysis, both at the population level and for the first time at the single-cell level, that differentiated cells express genes characteristic for the development of ANE, in particular for the development of the future forebrain. Stem Cells2012;30:1875–1884