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Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture

Nature Biotechnology volume 21, pages 183–186 (2003)Cite this article

Abstract

Mouse embryonic stem (ES) cells are competent for production of all fetal and adult cell types1. However, the utility of ES cells as a developmental model or as a source of defined cell populations for pharmaceutical screening or transplantation is compromised because their differentiation in vitro is poorly controlled2. Specification of primary lineages is not understood and consequently differentiation protocols are empirical, yielding variable and heterogeneous outcomes. Here we report that neither multicellular aggregation3,4 nor coculture5 is necessary for ES cells to commit efficiently to a neural fate. In adherent monoculture, elimination of inductive signals for alternative fates is sufficient for ES cells to develop into neural precursors. This process is not a simple default pathway, however, but requires autocrine fibroblast growth factor (FGF). Using flow cytometry quantitation and recording of individual colonies, we establish that the bulk of ES cells undergo neural conversion. The neural precursors can be purified to homogeneity by fluorescence activated cell sorting (FACS) or drug selection. This system provides a platform for defining the molecular machinery of neural commitment and optimizing the efficiency of neuronal and glial cell production from pluripotent mammalian stem cells.

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Figure 1: Sox1-GFP reporter of neural specification.
Figure 2: Neuronal and glial differentiation in monolayer culture.
Figure 3: Visualization of neural determination of ES cells.
Figure 4: Effects of growth factors and antagonists on neural determination.

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Acknowledgements

We thank John Heath for the DN-FGFR construct. We are grateful to Steve le Moenic for flow cytometry support and to Alexander Medvinsky and Josh Brickman for comments on the manuscript. This research is supported by the International Human Frontiers Science Program Organisation, and by the Medical Research Council and the Biotechnology and Biological Sciences Research Council of the United Kingdom.

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  1. Qi-Long Ying and Marios Stavridis: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Stem Cell Research, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh, EH9 3JQ, United Kingdom

    Qi-Long Ying, Marios Stavridis, Dean Griffiths, Meng Li & Austin Smith

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  1. Qi-Long Ying
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Correspondence to Austin Smith.

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A.S. is a scientific advisor to Stem Cell Sciences Ltd. and holds nonvoting equity in the company. Stem Cell Sciences funds research in the laboratory and has patents granted and pending on technology used in this manuscript.

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Ying, QL., Stavridis, M., Griffiths, D. et al. Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. Nat Biotechnol 21, 183–186 (2003). https://doi.org/10.1038/nbt780

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